Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion

Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin d...
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Autor*in:	Tobias Busche [verfasserIn] Konstantinos C. Tsolis [verfasserIn] Joachim Koepff [verfasserIn] Yuriy Rebets [verfasserIn] Christian Rückert [verfasserIn] Mohamed B. Hamed [verfasserIn] Arne Bleidt [verfasserIn] Wolfgang Wiechert [verfasserIn] Mariia Lopatniuk [verfasserIn] Ahmed Yousra [verfasserIn] Jozef Anné [verfasserIn] Spyridoula Karamanou [verfasserIn] Marco Oldiges [verfasserIn] Jörn Kalinowski [verfasserIn] Andriy Luzhetskyy [verfasserIn] Anastassios Economou [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2018

Schlagwörter:	proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion

Übergeordnetes Werk:	In: Frontiers in Microbiology - Frontiers Media S.A., 2011, 9(2018)
Übergeordnetes Werk:	volume:9 ; year:2018

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fmicb.2018.01174

Katalog-ID:	DOAJ060712694

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allfields	10.3389/fmicb.2018.01174 doi (DE-627)DOAJ060712694 (DE-599)DOAJe03a6f18fcd74f79ad14fb8e8b2d328a DE-627 ger DE-627 rakwb eng QR1-502 Tobias Busche verfasserin aut Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process. proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology Tobias Busche verfasserin aut Konstantinos C. Tsolis verfasserin aut Joachim Koepff verfasserin aut Yuriy Rebets verfasserin aut Yuriy Rebets verfasserin aut Christian Rückert verfasserin aut Mohamed B. Hamed verfasserin aut Mohamed B. Hamed verfasserin aut Arne Bleidt verfasserin aut Wolfgang Wiechert verfasserin aut Mariia Lopatniuk verfasserin aut Ahmed Yousra verfasserin aut Jozef Anné verfasserin aut Spyridoula Karamanou verfasserin aut Marco Oldiges verfasserin aut Marco Oldiges verfasserin aut Jörn Kalinowski verfasserin aut Andriy Luzhetskyy verfasserin aut Anastassios Economou verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 9(2018) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:9 year:2018 https://doi.org/10.3389/fmicb.2018.01174 kostenfrei https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2018
spelling	10.3389/fmicb.2018.01174 doi (DE-627)DOAJ060712694 (DE-599)DOAJe03a6f18fcd74f79ad14fb8e8b2d328a DE-627 ger DE-627 rakwb eng QR1-502 Tobias Busche verfasserin aut Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process. proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology Tobias Busche verfasserin aut Konstantinos C. Tsolis verfasserin aut Joachim Koepff verfasserin aut Yuriy Rebets verfasserin aut Yuriy Rebets verfasserin aut Christian Rückert verfasserin aut Mohamed B. Hamed verfasserin aut Mohamed B. Hamed verfasserin aut Arne Bleidt verfasserin aut Wolfgang Wiechert verfasserin aut Mariia Lopatniuk verfasserin aut Ahmed Yousra verfasserin aut Jozef Anné verfasserin aut Spyridoula Karamanou verfasserin aut Marco Oldiges verfasserin aut Marco Oldiges verfasserin aut Jörn Kalinowski verfasserin aut Andriy Luzhetskyy verfasserin aut Anastassios Economou verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 9(2018) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:9 year:2018 https://doi.org/10.3389/fmicb.2018.01174 kostenfrei https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2018
allfields_unstemmed	10.3389/fmicb.2018.01174 doi (DE-627)DOAJ060712694 (DE-599)DOAJe03a6f18fcd74f79ad14fb8e8b2d328a DE-627 ger DE-627 rakwb eng QR1-502 Tobias Busche verfasserin aut Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process. proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology Tobias Busche verfasserin aut Konstantinos C. Tsolis verfasserin aut Joachim Koepff verfasserin aut Yuriy Rebets verfasserin aut Yuriy Rebets verfasserin aut Christian Rückert verfasserin aut Mohamed B. Hamed verfasserin aut Mohamed B. Hamed verfasserin aut Arne Bleidt verfasserin aut Wolfgang Wiechert verfasserin aut Mariia Lopatniuk verfasserin aut Ahmed Yousra verfasserin aut Jozef Anné verfasserin aut Spyridoula Karamanou verfasserin aut Marco Oldiges verfasserin aut Marco Oldiges verfasserin aut Jörn Kalinowski verfasserin aut Andriy Luzhetskyy verfasserin aut Anastassios Economou verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 9(2018) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:9 year:2018 https://doi.org/10.3389/fmicb.2018.01174 kostenfrei https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2018
allfieldsGer	10.3389/fmicb.2018.01174 doi (DE-627)DOAJ060712694 (DE-599)DOAJe03a6f18fcd74f79ad14fb8e8b2d328a DE-627 ger DE-627 rakwb eng QR1-502 Tobias Busche verfasserin aut Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process. proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology Tobias Busche verfasserin aut Konstantinos C. Tsolis verfasserin aut Joachim Koepff verfasserin aut Yuriy Rebets verfasserin aut Yuriy Rebets verfasserin aut Christian Rückert verfasserin aut Mohamed B. Hamed verfasserin aut Mohamed B. Hamed verfasserin aut Arne Bleidt verfasserin aut Wolfgang Wiechert verfasserin aut Mariia Lopatniuk verfasserin aut Ahmed Yousra verfasserin aut Jozef Anné verfasserin aut Spyridoula Karamanou verfasserin aut Marco Oldiges verfasserin aut Marco Oldiges verfasserin aut Jörn Kalinowski verfasserin aut Andriy Luzhetskyy verfasserin aut Anastassios Economou verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 9(2018) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:9 year:2018 https://doi.org/10.3389/fmicb.2018.01174 kostenfrei https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2018
allfieldsSound	10.3389/fmicb.2018.01174 doi (DE-627)DOAJ060712694 (DE-599)DOAJe03a6f18fcd74f79ad14fb8e8b2d328a DE-627 ger DE-627 rakwb eng QR1-502 Tobias Busche verfasserin aut Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process. proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology Tobias Busche verfasserin aut Konstantinos C. Tsolis verfasserin aut Joachim Koepff verfasserin aut Yuriy Rebets verfasserin aut Yuriy Rebets verfasserin aut Christian Rückert verfasserin aut Mohamed B. Hamed verfasserin aut Mohamed B. Hamed verfasserin aut Arne Bleidt verfasserin aut Wolfgang Wiechert verfasserin aut Mariia Lopatniuk verfasserin aut Ahmed Yousra verfasserin aut Jozef Anné verfasserin aut Spyridoula Karamanou verfasserin aut Marco Oldiges verfasserin aut Marco Oldiges verfasserin aut Jörn Kalinowski verfasserin aut Andriy Luzhetskyy verfasserin aut Anastassios Economou verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 9(2018) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:9 year:2018 https://doi.org/10.3389/fmicb.2018.01174 kostenfrei https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2018
language	English
source	In Frontiers in Microbiology 9(2018) volume:9 year:2018
sourceStr	In Frontiers in Microbiology 9(2018) volume:9 year:2018
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion Microbiology
isfreeaccess_bool	true
container_title	Frontiers in Microbiology
authorswithroles_txt_mv	Tobias Busche @@aut@@ Konstantinos C. Tsolis @@aut@@ Joachim Koepff @@aut@@ Yuriy Rebets @@aut@@ Christian Rückert @@aut@@ Mohamed B. Hamed @@aut@@ Arne Bleidt @@aut@@ Wolfgang Wiechert @@aut@@ Mariia Lopatniuk @@aut@@ Ahmed Yousra @@aut@@ Jozef Anné @@aut@@ Spyridoula Karamanou @@aut@@ Marco Oldiges @@aut@@ Jörn Kalinowski @@aut@@ Andriy Luzhetskyy @@aut@@ Anastassios Economou @@aut@@
publishDateDaySort_date	2018-01-01T00:00:00Z
hierarchy_top_id	642889384
id	DOAJ060712694
language_de	englisch
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callnumber-first	Q - Science
author	Tobias Busche
spellingShingle	Tobias Busche misc QR1-502 misc proteases misc expression levels misc mRFP misc multi-omics misc Streptomyces lividans misc protein secretion misc Microbiology Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion
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topic_title	QR1-502 Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion proteases expression levels mRFP multi-omics Streptomyces lividans protein secretion
topic	misc QR1-502 misc proteases misc expression levels misc mRFP misc multi-omics misc Streptomyces lividans misc protein secretion misc Microbiology
topic_unstemmed	misc QR1-502 misc proteases misc expression levels misc mRFP misc multi-omics misc Streptomyces lividans misc protein secretion misc Microbiology
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title	Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion
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title_full	Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion
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author_browse	Tobias Busche Konstantinos C. Tsolis Joachim Koepff Yuriy Rebets Christian Rückert Mohamed B. Hamed Arne Bleidt Wolfgang Wiechert Mariia Lopatniuk Ahmed Yousra Jozef Anné Spyridoula Karamanou Marco Oldiges Jörn Kalinowski Andriy Luzhetskyy Anastassios Economou
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title_sort	multi-omics and targeted approaches to determine the role of cellular proteases in streptomyces protein secretion
callnumber	QR1-502
title_auth	Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion
abstract	Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process.
abstractGer	Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process.
abstract_unstemmed	Gram-positive Streptomyces bacteria are profuse secretors of polypeptides using complex, yet unknown mechanisms. Many of their secretory proteins are proteases that play important roles in the acquisition of amino acids from the environment. Other proteases regulate cellular proteostasis. To begin dissecting the possible role of proteases in Streptomyces secretion, we applied a multi-omics approach. We probed the role of the 190 proteases of Streptomyces lividans strain TK24 in protein secretion in defined media at different stages of growth. Transcriptomics analysis revealed transcripts for 93% of these proteases and identified that 41 of them showed high abundance. Proteomics analysis identified 57 membrane-embedded or secreted proteases with variations in their abundance. We focused on 17 of these proteases and putative inhibitors and generated strains deleted of their genes. These were characterized in terms of their fitness, transcriptome and secretome changes. In addition, we performed a targeted analysis in deletion strains that also carried a secretion competent mRFP. One strain, carrying a deletion of the gene for the regulatory protease FtsH, showed significant global changes in overall transcription and enhanced secretome and secreted mRFP levels. These data provide a first multi-omics effort to characterize the complex regulatory mechanisms of protein secretion in Streptomyces lividans and lay the foundations for future rational manipulation of this process.
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title_short	Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion
url	https://doi.org/10.3389/fmicb.2018.01174 https://doaj.org/article/e03a6f18fcd74f79ad14fb8e8b2d328a https://www.frontiersin.org/article/10.3389/fmicb.2018.01174/full https://doaj.org/toc/1664-302X
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up_date	2024-07-03T16:36:08.443Z
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