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Title: Cell-to-cell interaction requires optimal positioning of a pilus tip adhesin modulated by gram-positive transpeptidase enzymes

Abstract

Assembly of pili on the gram-positive bacterial cell wall involves 2 conserved transpeptidase enzymes named sortases: One for polymerization of pilin subunits and another for anchoring pili to peptidoglycan. How this machine controls pilus length and whether pilus length is critical for cell-to-cell interactions remain unknown. We report here in Actinomyces oris, a key colonizer in the development of oral biofilms, that genetic disruption of its housekeeping sortase SrtA generates exceedingly long pili, catalyzed by its pilus-specific sortase SrtC2 that possesses both pilus polymerization and cell wall anchoring functions. Remarkably, the srtA-deficient mutant fails to mediate interspecies interactions, or coaggregation, even though the coaggregation factor CafA is present at the pilus tip. Furthermore, increasing ectopic expression of srtA in the mutant progressively shortens pilus length and restores coaggregation accordingly, while elevated levels of shaft pilins and SrtC2 produce long pili and block coaggregation by SrtA + bacteria. With structural studies, we uncovered 2 key structural elements in SrtA that partake in recognition of pilin substrates and regulate pilus length by inducing the capture and transfer of pilus polymers to the cell wall. Evidently, coaggregation requires proper positioning of the tip adhesin CafA via modulation of pilus length by the housekeepingmore » sortase SrtA.« less

Authors:
 [1];  [2];  [3];  [2];  [4];  [2];  [3];  [5];  [6];  [7]
  1. Univ. of California, Los Angeles, CA (United States). Div. of Oral Biology and Medicine
  2. Univ. of Texas Health Science Center, Houston, TX (United States). Dept. of Microbiology & Molecular Genetics
  3. Univ. of Chicago, Chicago, IL (United States). Center for Structural Genomics of Infectious Diseases, Consortium for Advanced Science and Engineering; Argonne National Lab. (ANL), Lemont, IL (United States). Structural Biology Center
  4. Yale School of Medicine, New Haven, CT (United States). Dept. of Microbial Pathogenesis
  5. Univ. of California, Los Angeles, CA (United States). Dept. of Chemistry and Biochemistry
  6. Univ. of Connecticut, Storrs, CT (United States). Dept. of Medicine, Neag Comprehensive Cancer Center
  7. Univ. of California, Los Angeles, CA (United States). Div. of Oral Biology and Medicine and Molecular Biology Inst.
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Institutes of Health (NIH), National Institute of Dental and Craniofacial Research; University of Texas Health Science Center at Houston; National Institutes of Health (NIH), National Institute of Allergy and Infectious Diseases (NIAID)
OSTI Identifier:
1575069
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 36; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; coaggregation; gram-positive bacteria; pilus assembly; pilus length; sortase

Citation Formats

Chang, Chungyu, Wu, Chenggang, Osipiuk, Jerzy, Siegel, Sara D., Zhu, Shiwei, Liu, Xiangan, Joachimiak, Andrzej, Clubb, Robert T., Das, Asis, and Ton-That, Hung. Cell-to-cell interaction requires optimal positioning of a pilus tip adhesin modulated by gram-positive transpeptidase enzymes. United States: N. p., 2019. Web. doi:10.1073/pnas.1907733116.
Chang, Chungyu, Wu, Chenggang, Osipiuk, Jerzy, Siegel, Sara D., Zhu, Shiwei, Liu, Xiangan, Joachimiak, Andrzej, Clubb, Robert T., Das, Asis, & Ton-That, Hung. Cell-to-cell interaction requires optimal positioning of a pilus tip adhesin modulated by gram-positive transpeptidase enzymes. United States. doi:10.1073/pnas.1907733116.
Chang, Chungyu, Wu, Chenggang, Osipiuk, Jerzy, Siegel, Sara D., Zhu, Shiwei, Liu, Xiangan, Joachimiak, Andrzej, Clubb, Robert T., Das, Asis, and Ton-That, Hung. Mon . "Cell-to-cell interaction requires optimal positioning of a pilus tip adhesin modulated by gram-positive transpeptidase enzymes". United States. doi:10.1073/pnas.1907733116.
@article{osti_1575069,
title = {Cell-to-cell interaction requires optimal positioning of a pilus tip adhesin modulated by gram-positive transpeptidase enzymes},
author = {Chang, Chungyu and Wu, Chenggang and Osipiuk, Jerzy and Siegel, Sara D. and Zhu, Shiwei and Liu, Xiangan and Joachimiak, Andrzej and Clubb, Robert T. and Das, Asis and Ton-That, Hung},
abstractNote = {Assembly of pili on the gram-positive bacterial cell wall involves 2 conserved transpeptidase enzymes named sortases: One for polymerization of pilin subunits and another for anchoring pili to peptidoglycan. How this machine controls pilus length and whether pilus length is critical for cell-to-cell interactions remain unknown. We report here in Actinomyces oris, a key colonizer in the development of oral biofilms, that genetic disruption of its housekeeping sortase SrtA generates exceedingly long pili, catalyzed by its pilus-specific sortase SrtC2 that possesses both pilus polymerization and cell wall anchoring functions. Remarkably, the srtA-deficient mutant fails to mediate interspecies interactions, or coaggregation, even though the coaggregation factor CafA is present at the pilus tip. Furthermore, increasing ectopic expression of srtA in the mutant progressively shortens pilus length and restores coaggregation accordingly, while elevated levels of shaft pilins and SrtC2 produce long pili and block coaggregation by SrtA+ bacteria. With structural studies, we uncovered 2 key structural elements in SrtA that partake in recognition of pilin substrates and regulate pilus length by inducing the capture and transfer of pilus polymers to the cell wall. Evidently, coaggregation requires proper positioning of the tip adhesin CafA via modulation of pilus length by the housekeeping sortase SrtA.},
doi = {10.1073/pnas.1907733116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 36,
volume = 116,
place = {United States},
year = {2019},
month = {8}
}

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