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A CRISPR with Roles in Myxococcus xanthus Development and Exopolysaccharide Production

dc.contributorVirginia Techen
dc.contributor.authorWallace, Regina A.en
dc.contributor.authorBlack, Wesley P.en
dc.contributor.authorYang, Xianshuangen
dc.contributor.authorYang, Zhaominen
dc.contributor.departmentBiological Sciencesen
dc.date.accessed2015-10-01en
dc.date.accessioned2015-11-28T21:53:46Zen
dc.date.available2015-11-28T21:53:46Zen
dc.date.issued2014-09-08en
dc.description.abstractThe Gram-negative soil bacterium Myxococcus xanthus utilizes its social (S) gliding motility to move on surfaces during its vegetative and developmental cycles. It is known that S motility requires the type IV pilus (T4P) and the exopolysaccharide (EPS) to function. The T4P is the S motility motor, and it powers cell movement by retraction. As the key regulator of the S motor, EPS is proposed to be the anchor and trigger for T4P retraction. The production of EPS is regulated in turn by the T4P in M. xanthus, and T4P(-) mutants are S- and EPS-. In this study, a Delta pilA strain (T4P(-) and EPS-) was mutagenized by a transposon and screened for EPS- mutants. A pilA suppressor isolated as such harbored an insertion in the 3rd clustered regularly interspaced short palindromic repeat (CRISPR3) in M. xanthus. Evidence indicates that this transposon insertion, designated CRISPR3(star), is a gain-of-function (GOF) mutation. Moreover, CRISPR3(star) eliminated developmental aggregation in both the wild-type and the pilA mutant backgrounds. Upstream of CRISPR3 are genes encoding the repeat-associated mysterious proteins (RAMPs). These RAMP genes are indispensable for CRISPR3(star) to affect development and EPS in M. xanthus. Analysis by reverse transcription (RT)-PCR suggested that CRISPR3(star) led to an increase in the processing of the RNA transcribed from CRISPR3. We propose that certain CRISPR3 transcripts, once expressed and processed, target genes critical for M. xanthus fruiting body development and EPS production in a RAMP-dependent manner.en
dc.description.sponsorshipNational Science Foundationen
dc.description.sponsorshipMCB 1239889en
dc.description.sponsorshipMCB-1417726en
dc.description.sponsorshipNational Institutes of Healthen
dc.description.sponsorshipGM071601en
dc.description.sponsorshipVirginia Tech. Fralin Life Science Instituteen
dc.description.sponsorshipVirginia Tech. Post Baccalaureate Research and Education Program (PREP)en
dc.description.sponsorshipVirginia Tech. Multicultural Academic Opportunities Program (MAOP)en
dc.format.mimetypeapplication/pdfen
dc.identifier.citationWallace, Regina A., et al. (2014). A CRISPR with Roles in Myxococcus xanthus Development and Exopolysaccharide Production. Journal of Bacteriology, 196(23), 4036-4043. doi:10.1128/jb.02035-14en
dc.identifier.doihttps://doi.org/10.1128/jb.02035-14en
dc.identifier.issn0021-9193en
dc.identifier.urihttp://hdl.handle.net/10919/64209en
dc.identifier.urlhttp://jb.asm.org/content/196/23/4036en
dc.language.isoen_USen
dc.publisherAmerican Society for Microbiologyen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.titleA CRISPR with Roles in Myxococcus xanthus Development and Exopolysaccharide Productionen
dc.title.serialJournal of Bacteriologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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