TY - JOUR
T1 - Recognition of extracellular DNA by type IV pili promotes biofilm formation by Clostridioides difficile
AU - Ronish, Leslie A.
AU - Sidner, Ben
AU - Yu, Yafan
AU - Piepenbrink, Kurt H.
N1 - Funding Information:
We thank the staff at Argonne National Labs Advanced Photon Source General Medical Sciences and Cancer Institutes of Structural Biology Facility (GM/CA) and the staff at Stanford Synchrotron Radiation Lightsource beam line 12-2 for technical assistance with X-ray data collection. We also acknowledge technical assistance by Troy Syed in protein expression and purification and the staff of the UNL Microscopy Core for scanning electron microscopy and CLSM imaging. L. A. R. investigation; L. A. R. formal analysis; L. A. R. writing–original draft; B. S. validation; Y. Y. methodology; K. H. P. supervision; K. H. P. funding acquisition; K. H. P. writing–review and editing. This work was supported by National Institutes of Health grants K22-AI123467 (to K. H. P.) and P20-GM113126 (K. H. P. was a young investigator through the Nebraska Center for Integrated Biomolecular Communication). Additionally, L. A. R. was supported by T32-GM107001. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
This work was supported by National Institutes of Health grants K22-AI123467 (to K. H. P.) and P20-GM113126 (K. H. P. was a young investigator through the Nebraska Center for Integrated Biomolecular Communication). Additionally, L. A. R. was supported by T32-GM107001. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 The Authors
PY - 2022/10
Y1 - 2022/10
N2 - Clostridioides difficile is a Gram-positive bacillus, which is a frequent cause of gastrointestinal infections triggered by the depletion of the gut microbiome. Because of the frequent recurrence of these infections after antibiotic treatment, mechanisms of C. difficile persistence and recurrence, including biofilm formation, are of increasing interest. Previously, our group and others found that type IV pili, filamentous helical appendages polymerized from protein subunits, promoted microcolony and biofilm formation in C. difficile. In Gram-negative bacteria, the ability of type IV pili to mediate bacterial self-association has been explained through interactions between the pili of adjacent cells, but type IV pili from several Gram-negative species are also required for natural competence through DNA uptake. Here, we report the ability of two C. difficile pilin subunits, PilJ and PilW, to bind to DNA in vitro, as well as the defects in biofilm formation in the pilJ and pilW gene-interruption mutants. Additionally, we have resolved the X-ray crystal structure of PilW, which we use to model possible structural mechanisms for the formation of C. difficile biofilm through interactions between type IV pili and the DNA of the extracellular matrix. Taken together, our results provide further insight into the relationship between type IV pilus function and biofilm formation in C. difficile and, more broadly, suggest that DNA recognition by type IV pili and related structures may have functional importance beyond DNA uptake for natural competence.
AB - Clostridioides difficile is a Gram-positive bacillus, which is a frequent cause of gastrointestinal infections triggered by the depletion of the gut microbiome. Because of the frequent recurrence of these infections after antibiotic treatment, mechanisms of C. difficile persistence and recurrence, including biofilm formation, are of increasing interest. Previously, our group and others found that type IV pili, filamentous helical appendages polymerized from protein subunits, promoted microcolony and biofilm formation in C. difficile. In Gram-negative bacteria, the ability of type IV pili to mediate bacterial self-association has been explained through interactions between the pili of adjacent cells, but type IV pili from several Gram-negative species are also required for natural competence through DNA uptake. Here, we report the ability of two C. difficile pilin subunits, PilJ and PilW, to bind to DNA in vitro, as well as the defects in biofilm formation in the pilJ and pilW gene-interruption mutants. Additionally, we have resolved the X-ray crystal structure of PilW, which we use to model possible structural mechanisms for the formation of C. difficile biofilm through interactions between type IV pili and the DNA of the extracellular matrix. Taken together, our results provide further insight into the relationship between type IV pilus function and biofilm formation in C. difficile and, more broadly, suggest that DNA recognition by type IV pili and related structures may have functional importance beyond DNA uptake for natural competence.
KW - C. difficile
KW - X-ray crystallography
KW - adhesion
KW - bacterial biofilm
KW - extracellular matrix
KW - protein DNA interactions
KW - structural biology
KW - surface motility
KW - type IV pili
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UR - http://www.scopus.com/inward/citedby.url?scp=85139591150&partnerID=8YFLogxK
U2 - 10.1016/j.jbc.2022.102449
DO - 10.1016/j.jbc.2022.102449
M3 - Article
C2 - 36064001
AN - SCOPUS:85139591150
SN - 0021-9258
VL - 298
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 10
M1 - 102449
ER -