A central role for the transcriptional regulator VtlR in small RNA-mediated gene regulation in Agrobacterium tumefaciens

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dc.contributor.authorBudnick, James A.en
dc.contributor.authorSheehan, Lauren M.en
dc.contributor.authorGinder, Miranda. J.en
dc.contributor.authorFailor, Kevin C.en
dc.contributor.authorPerkowski, Julia. M.en
dc.contributor.authorPinto, John. F.en
dc.contributor.authorKohl, Kirsten A.en
dc.contributor.authorKang, Linen
dc.contributor.authorMichalak, Pawelen
dc.contributor.authorLuo, Lien
dc.contributor.authorHeindl, Jason E.en
dc.contributor.authorCaswell, Clayton C.en
dc.contributor.departmentBiomedical Sciences and Pathobiologyen
dc.date.accessioned2021-02-08T20:02:36Zen
dc.date.available2021-02-08T20:02:36Zen
dc.date.issued2020-09-11en
dc.description.abstractLysR-type transcriptional regulators (LTTRs) are the most common type of transcriptional regulators in prokaryotes and function by altering gene expression in response to environmental stimuli. In the class Alphaproteobacteria, a conserved LTTR named VtlR is critical to the establishment of host-microbe interactions. In the mammalian pathogen Brucella abortus, VtlR is required for full virulence in a mouse model of infection, and VtlR activates the expression of abcR2, which encodes a small regulatory RNA (sRNA). In the plant symbiont Sinorhizobium meliloti, the ortholog of VtlR, named LsrB, is involved in the symbiosis of the bacterium with alfalfa. Agrobacterium tumefaciens is a close relative of both B. abortus and S. meliloti, and this bacterium is the causative agent of crown gall disease in plants. In the present study, we demonstrate that VtlR is involved in the ability of A. tumefaciens to grow appropriately in artificial medium, and an A. tumefaciens vtlR deletion strain is defective in motility, biofilm formation, and tumorigenesis of potato discs. RNA-sequencing analyses revealed that more than 250 genes are dysregulated in the vtlR strain, and importantly, VtlR directly controls the expression of three sRNAs in A. tumefaciens. Taken together, these data support a model in which VtlR indirectly regulates hundreds of genes via manipulation of sRNA pathways in A. tumefaciens, and moreover, while the VtlR/LsrB protein is present and structurally conserved in many members of the Alphaproteobacteria, the VtlR/LsrB regulatory circuitry has diverged in order to accommodate the unique environmental niche of each organism.en
dc.description.notesWe would like to thank Tristan Stoyanof for assistance with EMSAs and annotation of RNA-seq data. We would like to thank Gilda Bocco, Rachel Kesselring, and Hayleigh Monroe for assistance with swimming motility and biofilm formation assays. This work was funded by a One Health Grant from the Edward Via College of Osteopathic Medicine (CCC), Grant H1804 from the W.W. Smith Charitable Trust (JEH), and Grant 1R15GM135933-01 from the NIH/NIGMS (JEH), and we are very grateful for this financial support. MJG was supported by a Melvin Firman Undergraduate Summer Research Grant and an Arno Viehover Undergraduate Tuition Scholarship, both from the University of the Sciences in Philadelphia.en
dc.description.sponsorshipOne Health Grant from the Edward Via College of Osteopathic Medicine; W.W. Smith Charitable Trust [H1804]; NIH/NIGMSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [1R15GM135933-01]; Melvin Firman Undergraduate Summer Research Grant from the University of the Sciences in Philadelphia; Arno Viehover Undergraduate Tuition Scholarship from the University of the Sciences in Philadelphiaen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1038/s41598-020-72117-0en
dc.identifier.issn2045-2322en
dc.identifier.issue1en
dc.identifier.other14968en
dc.identifier.pmid32917931en
dc.identifier.urihttp://hdl.handle.net/10919/102310en
dc.identifier.volume10en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.titleA central role for the transcriptional regulator VtlR in small RNA-mediated gene regulation in Agrobacterium tumefaciensen
dc.title.serialScientific Reportsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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