Suppressor Mutations in Type II Secretion Mutants of Vibrio cholerae: Inactivation of the VesC Protease

Rule, Chelsea S.; Park, Young-Jun; Delarosa, Jaclyn R.; Turley, Stewart; Hol, Wim G.  J.; McColm, Sarah; Gura, Colby; DiMaio, Frank; Korotkov, Konstantin V.; Sandkvist, Maria

doi:10.1128/mSphere.01125-20

Suppressor Mutations in Type II Secretion Mutants of Vibrio cholerae: Inactivation of the VesC Protease

Journal Article · Tue Dec 15 23:00:00 EST 2020 · mSphere

DOI:https://doi.org/10.1128/mSphere.01125-20· OSTI ID:1816352

Rule, Chelsea S. ^[1]; Park, Young-Jun ^[2]; Delarosa, Jaclyn R. ^[2]; Turley, Stewart ^[2]; Hol, Wim G. J. ^[3]; McColm, Sarah ^[4]; Gura, Colby ^[4]; DiMaio, Frank ^[2]; ^[5]; Sandkvist, Maria ^[4]

Univ. of Michigan, Ann Arbor, MI (United States). Dept. of Microbiology and Immunology; OSTI
Univ. of Washington, Seattle, WA (United States). Dept. of Biochemistry
Department of Biochemistry, University of Washington, Seattle, Washington, USA
Univ. of Michigan, Ann Arbor, MI (United States). Dept. of Microbiology and Immunology
Univ. of Washington, Seattle, WA (United States). Dept. of Biochemistry; Univ. of Kentucky, Lexington, KY (United States). Dept. of Molecular and Cellular Biochemistry

The type II secretion system (T2SS) is a conserved transport pathway responsible for the secretion of a range of virulence factors by many pathogens, including Vibrio cholerae. Disruption of the T2SS genes in V. cholerae results in loss of secretion, changes in cell envelope function, and growth defects. While T2SS mutants are viable, high-throughput genomic analyses have listed these genes among essential genes. To investigate whether secondary mutations arise as a consequence of T2SS inactivation, we sequenced the genomes of six V. cholerae T2SS mutants with deletions or insertions in either the epsG, epsL, or epsM genes and identified secondary mutations in all mutants. Two of the six T2SS mutants contain distinct mutations in the gene encoding the T2SS-secreted protease VesC. Other mutations were found in genes coding for V. cholerae cell envelope proteins. Subsequent sequence analysis of the vesC gene in 92 additional T2SS mutant isolates identified another 19 unique mutations including insertions or deletions, sequence duplications, and single-nucleotide changes resulting in amino acid substitutions in the VesC protein. Analysis of VesC variants and the X-ray crystallographic structure of wild-type VesC suggested that all mutations lead to loss of VesC production and/or function. One possible mechanism by which V. cholerae T2SS mutagenesis can be tolerated is through selection of vesC-inactivating mutations, which may, in part, suppress cell envelope damage, establishing permissive conditions for the disruption of the T2SS. Other mutations may have been acquired in genes encoding essential cell envelope proteins to prevent proteolysis by VesC.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Institute of Allergy and Infectious Diseases; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1816352

Journal Information:: mSphere, Journal Name: mSphere Journal Issue: 6 Vol. 5; ISSN 2379-5042

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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serine protease
suppressor
type II secretion system

Suppressor Mutations in Type II Secretion Mutants of Vibrio cholerae: Inactivation of the VesC Protease

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