Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8
Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or und...
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| Autor*in: |
Pandey, Himanshu [verfasserIn] Singh, Sudhir Kumar [verfasserIn] Sadan, Mayan [verfasserIn] Popov, Mary [verfasserIn] Singh, Meenakshi [verfasserIn] Davidov, Geula [verfasserIn] Inagaki, Sayaka [verfasserIn] Al-Bassam, Jawdat [verfasserIn] Zarivach, Raz [verfasserIn] Rosenfeld, Steven S. [verfasserIn] Gheber, Larisa [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2021 |
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| Schlagwörter: |
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| Anmerkung: |
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 |
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| Übergeordnetes Werk: |
Enthalten in: Cellular and molecular life sciences - Cham (ZG) : Springer International Publishing AG, 1997, 78(2021), 16 vom: 17. Juli, Seite 6051-6068 |
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| Übergeordnetes Werk: |
volume:78 ; year:2021 ; number:16 ; day:17 ; month:07 ; pages:6051-6068 |
| Links: |
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| DOI / URN: |
10.1007/s00018-021-03891-x |
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| Katalog-ID: |
SPR044674368 |
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| 100 | 1 | |a Pandey, Himanshu |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
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| 500 | |a © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 | ||
| 520 | |a Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. | ||
| 650 | 4 | |a Kinesin-5 |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Cin8 |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Motor–microtubule interaction |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Context-dependent directionality reversal |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Bi-directional motility |7 (dpeaa)DE-He213 | |
| 700 | 1 | |a Singh, Sudhir Kumar |e verfasserin |4 aut | |
| 700 | 1 | |a Sadan, Mayan |e verfasserin |4 aut | |
| 700 | 1 | |a Popov, Mary |e verfasserin |4 aut | |
| 700 | 1 | |a Singh, Meenakshi |e verfasserin |4 aut | |
| 700 | 1 | |a Davidov, Geula |e verfasserin |4 aut | |
| 700 | 1 | |a Inagaki, Sayaka |e verfasserin |4 aut | |
| 700 | 1 | |a Al-Bassam, Jawdat |e verfasserin |4 aut | |
| 700 | 1 | |a Zarivach, Raz |e verfasserin |4 aut | |
| 700 | 1 | |a Rosenfeld, Steven S. |e verfasserin |4 aut | |
| 700 | 1 | |a Gheber, Larisa |e verfasserin |4 aut | |
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10.1007/s00018-021-03891-x doi (DE-627)SPR044674368 (SPR)s00018-021-03891-x-e DE-627 ger DE-627 rakwb eng 570 610 ASE 42.15 bkl Pandey, Himanshu verfasserin aut Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 Singh, Sudhir Kumar verfasserin aut Sadan, Mayan verfasserin aut Popov, Mary verfasserin aut Singh, Meenakshi verfasserin aut Davidov, Geula verfasserin aut Inagaki, Sayaka verfasserin aut Al-Bassam, Jawdat verfasserin aut Zarivach, Raz verfasserin aut Rosenfeld, Steven S. verfasserin aut Gheber, Larisa verfasserin aut Enthalten in Cellular and molecular life sciences Cham (ZG) : Springer International Publishing AG, 1997 78(2021), 16 vom: 17. Juli, Seite 6051-6068 (DE-627)253390524 (DE-600)1458497-9 1420-9071 nnns volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 https://dx.doi.org/10.1007/s00018-021-03891-x lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_165 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.15 ASE AR 78 2021 16 17 07 6051-6068 |
| spelling |
10.1007/s00018-021-03891-x doi (DE-627)SPR044674368 (SPR)s00018-021-03891-x-e DE-627 ger DE-627 rakwb eng 570 610 ASE 42.15 bkl Pandey, Himanshu verfasserin aut Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 Singh, Sudhir Kumar verfasserin aut Sadan, Mayan verfasserin aut Popov, Mary verfasserin aut Singh, Meenakshi verfasserin aut Davidov, Geula verfasserin aut Inagaki, Sayaka verfasserin aut Al-Bassam, Jawdat verfasserin aut Zarivach, Raz verfasserin aut Rosenfeld, Steven S. verfasserin aut Gheber, Larisa verfasserin aut Enthalten in Cellular and molecular life sciences Cham (ZG) : Springer International Publishing AG, 1997 78(2021), 16 vom: 17. Juli, Seite 6051-6068 (DE-627)253390524 (DE-600)1458497-9 1420-9071 nnns volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 https://dx.doi.org/10.1007/s00018-021-03891-x lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_165 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.15 ASE AR 78 2021 16 17 07 6051-6068 |
| allfields_unstemmed |
10.1007/s00018-021-03891-x doi (DE-627)SPR044674368 (SPR)s00018-021-03891-x-e DE-627 ger DE-627 rakwb eng 570 610 ASE 42.15 bkl Pandey, Himanshu verfasserin aut Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 Singh, Sudhir Kumar verfasserin aut Sadan, Mayan verfasserin aut Popov, Mary verfasserin aut Singh, Meenakshi verfasserin aut Davidov, Geula verfasserin aut Inagaki, Sayaka verfasserin aut Al-Bassam, Jawdat verfasserin aut Zarivach, Raz verfasserin aut Rosenfeld, Steven S. verfasserin aut Gheber, Larisa verfasserin aut Enthalten in Cellular and molecular life sciences Cham (ZG) : Springer International Publishing AG, 1997 78(2021), 16 vom: 17. Juli, Seite 6051-6068 (DE-627)253390524 (DE-600)1458497-9 1420-9071 nnns volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 https://dx.doi.org/10.1007/s00018-021-03891-x lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_165 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.15 ASE AR 78 2021 16 17 07 6051-6068 |
| allfieldsGer |
10.1007/s00018-021-03891-x doi (DE-627)SPR044674368 (SPR)s00018-021-03891-x-e DE-627 ger DE-627 rakwb eng 570 610 ASE 42.15 bkl Pandey, Himanshu verfasserin aut Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 Singh, Sudhir Kumar verfasserin aut Sadan, Mayan verfasserin aut Popov, Mary verfasserin aut Singh, Meenakshi verfasserin aut Davidov, Geula verfasserin aut Inagaki, Sayaka verfasserin aut Al-Bassam, Jawdat verfasserin aut Zarivach, Raz verfasserin aut Rosenfeld, Steven S. verfasserin aut Gheber, Larisa verfasserin aut Enthalten in Cellular and molecular life sciences Cham (ZG) : Springer International Publishing AG, 1997 78(2021), 16 vom: 17. Juli, Seite 6051-6068 (DE-627)253390524 (DE-600)1458497-9 1420-9071 nnns volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 https://dx.doi.org/10.1007/s00018-021-03891-x lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_165 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.15 ASE AR 78 2021 16 17 07 6051-6068 |
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10.1007/s00018-021-03891-x doi (DE-627)SPR044674368 (SPR)s00018-021-03891-x-e DE-627 ger DE-627 rakwb eng 570 610 ASE 42.15 bkl Pandey, Himanshu verfasserin aut Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 Singh, Sudhir Kumar verfasserin aut Sadan, Mayan verfasserin aut Popov, Mary verfasserin aut Singh, Meenakshi verfasserin aut Davidov, Geula verfasserin aut Inagaki, Sayaka verfasserin aut Al-Bassam, Jawdat verfasserin aut Zarivach, Raz verfasserin aut Rosenfeld, Steven S. verfasserin aut Gheber, Larisa verfasserin aut Enthalten in Cellular and molecular life sciences Cham (ZG) : Springer International Publishing AG, 1997 78(2021), 16 vom: 17. Juli, Seite 6051-6068 (DE-627)253390524 (DE-600)1458497-9 1420-9071 nnns volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 https://dx.doi.org/10.1007/s00018-021-03891-x lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_165 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.15 ASE AR 78 2021 16 17 07 6051-6068 |
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Enthalten in Cellular and molecular life sciences 78(2021), 16 vom: 17. Juli, Seite 6051-6068 volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 |
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Enthalten in Cellular and molecular life sciences 78(2021), 16 vom: 17. Juli, Seite 6051-6068 volume:78 year:2021 number:16 day:17 month:07 pages:6051-6068 |
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Kinesin-5 Cin8 Motor–microtubule interaction Context-dependent directionality reversal Bi-directional motility |
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Pandey, Himanshu @@aut@@ Singh, Sudhir Kumar @@aut@@ Sadan, Mayan @@aut@@ Popov, Mary @@aut@@ Singh, Meenakshi @@aut@@ Davidov, Geula @@aut@@ Inagaki, Sayaka @@aut@@ Al-Bassam, Jawdat @@aut@@ Zarivach, Raz @@aut@@ Rosenfeld, Steven S. @@aut@@ Gheber, Larisa @@aut@@ |
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| author |
Pandey, Himanshu |
| spellingShingle |
Pandey, Himanshu ddc 570 bkl 42.15 misc Kinesin-5 misc Cin8 misc Motor–microtubule interaction misc Context-dependent directionality reversal misc Bi-directional motility Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
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570 610 ASE 42.15 bkl Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 Kinesin-5 (dpeaa)DE-He213 Cin8 (dpeaa)DE-He213 Motor–microtubule interaction (dpeaa)DE-He213 Context-dependent directionality reversal (dpeaa)DE-He213 Bi-directional motility (dpeaa)DE-He213 |
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ddc 570 bkl 42.15 misc Kinesin-5 misc Cin8 misc Motor–microtubule interaction misc Context-dependent directionality reversal misc Bi-directional motility |
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ddc 570 bkl 42.15 misc Kinesin-5 misc Cin8 misc Motor–microtubule interaction misc Context-dependent directionality reversal misc Bi-directional motility |
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ddc 570 bkl 42.15 misc Kinesin-5 misc Cin8 misc Motor–microtubule interaction misc Context-dependent directionality reversal misc Bi-directional motility |
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Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
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Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
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Pandey, Himanshu |
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Cellular and molecular life sciences |
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Pandey, Himanshu Singh, Sudhir Kumar Sadan, Mayan Popov, Mary Singh, Meenakshi Davidov, Geula Inagaki, Sayaka Al-Bassam, Jawdat Zarivach, Raz Rosenfeld, Steven S. Gheber, Larisa |
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Pandey, Himanshu |
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10.1007/s00018-021-03891-x |
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570 610 |
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verfasserin |
| title_sort |
flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 cin8 |
| title_auth |
Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
| abstract |
Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 |
| abstractGer |
Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 |
| abstract_unstemmed |
Abstract Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor–microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay. © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021. corrected publication 2021 |
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Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8 |
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Singh, Sudhir Kumar Sadan, Mayan Popov, Mary Singh, Meenakshi Davidov, Geula Inagaki, Sayaka Al-Bassam, Jawdat Zarivach, Raz Rosenfeld, Steven S. Gheber, Larisa |
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Singh, Sudhir Kumar Sadan, Mayan Popov, Mary Singh, Meenakshi Davidov, Geula Inagaki, Sayaka Al-Bassam, Jawdat Zarivach, Raz Rosenfeld, Steven S. Gheber, Larisa |
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10.1007/s00018-021-03891-x |
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