Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2
Four serine/threonine kinases are present in all mycobacteria: PknA, PknB, PknG and PknL. PknA and PknB are essential for growth and replication, PknG regulates metabolism, but little is known about PknL. Inactivation of pknL and adjacent regulator MSMEG_4242 in rough colony M. smegmatis mc2155 prod...
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Format: | Article |
Language: | English |
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Elsevier
2021-12-01
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Series: | The Cell Surface |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S246823302100013X |
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doaj-611ee645f9a642f8b53d9cbe636b8cf3 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Estalina Báez-Ramírez Luis Querales Carlos Andres Aranaga Gustavo López Elba Guerrero Laurent Kremer Séverine Carrère-Kremer Albertus Viljoen Mamadou Daffé Françoise Laval Stewart T. Cole Andrej Benjak Pedro Alzari Gwenaëlle André-Leroux William R. Jacobs, Jr. Catherine Vilcheze Howard E. Takiff |
spellingShingle |
Estalina Báez-Ramírez Luis Querales Carlos Andres Aranaga Gustavo López Elba Guerrero Laurent Kremer Séverine Carrère-Kremer Albertus Viljoen Mamadou Daffé Françoise Laval Stewart T. Cole Andrej Benjak Pedro Alzari Gwenaëlle André-Leroux William R. Jacobs, Jr. Catherine Vilcheze Howard E. Takiff Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 The Cell Surface Tuberculosis Kinase PknL Lsr2 Biofilms Mycobacterial envelope |
author_facet |
Estalina Báez-Ramírez Luis Querales Carlos Andres Aranaga Gustavo López Elba Guerrero Laurent Kremer Séverine Carrère-Kremer Albertus Viljoen Mamadou Daffé Françoise Laval Stewart T. Cole Andrej Benjak Pedro Alzari Gwenaëlle André-Leroux William R. Jacobs, Jr. Catherine Vilcheze Howard E. Takiff |
author_sort |
Estalina Báez-Ramírez |
title |
Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 |
title_short |
Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 |
title_full |
Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 |
title_fullStr |
Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 |
title_full_unstemmed |
Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2 |
title_sort |
elimination of pknl and msmeg_4242 in mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in lsr2 |
publisher |
Elsevier |
series |
The Cell Surface |
issn |
2468-2330 |
publishDate |
2021-12-01 |
description |
Four serine/threonine kinases are present in all mycobacteria: PknA, PknB, PknG and PknL. PknA and PknB are essential for growth and replication, PknG regulates metabolism, but little is known about PknL. Inactivation of pknL and adjacent regulator MSMEG_4242 in rough colony M. smegmatis mc2155 produced both smooth and rough colonies. Upon restreaking rough colonies, smooth colonies appeared at a frequency of ~ 1/250. Smooth mutants did not form biofilms, showed increased sliding motility and anomalous lipids on thin-layer chromatography, identified by mass spectrometry as lipooligosaccharides and perhaps also glycopeptidolipids. RNA-seq and Sanger sequencing revealed that all smooth mutants had inactivated lsr2 genes due to mutations and different IS1096 insertions. When complemented with lsr2, the colonies became rough, anomalous lipids disappeared and sliding motility decreased. Smooth mutants showed increased expression of IS1096 transposase TnpA and MSMEG_4727, which encodes a protein similar to PKS5. When MSMEG_4727 was deleted, smooth pknL/MSMEG_4242/lsr2 mutants reverted to rough, formed good biofilms, their motility decreased slightly and their anomalous lipids disappeared. Rough delpknL/del4242 mutants formed poor biofilms and showed decreased, aberrant sliding motility and both phenotypes were complemented with the two deleted genes. Inactivation of lsr2 changes colony morphology from rough to smooth, augments sliding motility and increases expression of MSMEG_4727 and other enzymes synthesizing lipooligosaccharides, apparently preventing biofilm formation. Similar morphological phase changes occur in other mycobacteria, likely reflecting environmental adaptations. PknL and MSMEG_4242 regulate lipid components of the outer cell envelope and their absence selects for lsr2 inactivation. A regulatory, phosphorylation cascade model is proposed. |
topic |
Tuberculosis Kinase PknL Lsr2 Biofilms Mycobacterial envelope |
url |
http://www.sciencedirect.com/science/article/pii/S246823302100013X |
work_keys_str_mv |
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doaj-611ee645f9a642f8b53d9cbe636b8cf32021-09-01T04:22:24ZengElsevierThe Cell Surface2468-23302021-12-017100060Elimination of PknL and MSMEG_4242 in Mycobacterium smegmatis alters the character of the outer cell envelope and selects for mutations in Lsr2Estalina Báez-Ramírez0Luis Querales1Carlos Andres Aranaga2Gustavo López3Elba Guerrero4Laurent Kremer5Séverine Carrère-Kremer6Albertus Viljoen7Mamadou Daffé8Françoise Laval9Stewart T. Cole10Andrej Benjak11Pedro Alzari12Gwenaëlle André-Leroux13William R. Jacobs, Jr.14Catherine Vilcheze15Howard E. Takiff16Laboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, VenezuelaLaboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, VenezuelaLaboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, VenezuelaLaboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, VenezuelaLaboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, VenezuelaInstitut de Recherche en Infectiologie de Montpellier, Centre National de la Recherche Scientifique UMR 9004, Université de Montpellier, Montpellier, FranceInstitut de Recherche en Infectiologie de Montpellier, Centre National de la Recherche Scientifique UMR 9004, Université de Montpellier, Montpellier, FranceInstitut de Recherche en Infectiologie de Montpellier, Centre National de la Recherche Scientifique UMR 9004, Université de Montpellier, Montpellier, FranceInstitut de Pharmacologie et de Biologie Structurale, Université de Toulouse, FranceInstitut de Pharmacologie et de Biologie Structurale, Université de Toulouse, FranceEPFL, Lauzanne, SwitzerlandEPFL, Lauzanne, SwitzerlandUnit of Structural Microbiology, Institut Pasteur, Paris, FranceUnit of Structural Microbiology, Institut Pasteur, Paris, FranceDepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USALaboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela; Shenzhen Nanshan CCDC, 7 Huaming Road, Nanshan, Shenzhen, China; Corresponding author at: Laboratorio de Genétic Molecular, CMBC, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela.Four serine/threonine kinases are present in all mycobacteria: PknA, PknB, PknG and PknL. PknA and PknB are essential for growth and replication, PknG regulates metabolism, but little is known about PknL. Inactivation of pknL and adjacent regulator MSMEG_4242 in rough colony M. smegmatis mc2155 produced both smooth and rough colonies. Upon restreaking rough colonies, smooth colonies appeared at a frequency of ~ 1/250. Smooth mutants did not form biofilms, showed increased sliding motility and anomalous lipids on thin-layer chromatography, identified by mass spectrometry as lipooligosaccharides and perhaps also glycopeptidolipids. RNA-seq and Sanger sequencing revealed that all smooth mutants had inactivated lsr2 genes due to mutations and different IS1096 insertions. When complemented with lsr2, the colonies became rough, anomalous lipids disappeared and sliding motility decreased. Smooth mutants showed increased expression of IS1096 transposase TnpA and MSMEG_4727, which encodes a protein similar to PKS5. When MSMEG_4727 was deleted, smooth pknL/MSMEG_4242/lsr2 mutants reverted to rough, formed good biofilms, their motility decreased slightly and their anomalous lipids disappeared. Rough delpknL/del4242 mutants formed poor biofilms and showed decreased, aberrant sliding motility and both phenotypes were complemented with the two deleted genes. Inactivation of lsr2 changes colony morphology from rough to smooth, augments sliding motility and increases expression of MSMEG_4727 and other enzymes synthesizing lipooligosaccharides, apparently preventing biofilm formation. Similar morphological phase changes occur in other mycobacteria, likely reflecting environmental adaptations. PknL and MSMEG_4242 regulate lipid components of the outer cell envelope and their absence selects for lsr2 inactivation. A regulatory, phosphorylation cascade model is proposed.http://www.sciencedirect.com/science/article/pii/S246823302100013XTuberculosisKinasePknLLsr2BiofilmsMycobacterial envelope |