Engineering a Conserved RNA Regulatory Protein Repurposes its Biological Function in Vivo

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dc.contributor.ORCID0000-0002-3768-6996 (Campbell, ZT)
dc.contributor.authorBhat, Vandita D.
dc.contributor.authorMcCann, Kathleen L.
dc.contributor.authorWang, Yeming
dc.contributor.authorFonseca, Dallas R.
dc.contributor.authorShukla, Tarjani
dc.contributor.authorAlexander, Jacqueline C.
dc.contributor.authorQiu, Chen
dc.contributor.authorWickens, Marv
dc.contributor.authorLo, Te-Wen
dc.contributor.authorHall, Traci M. Tanaka
dc.contributor.authorCampbell, Zachary T.
dc.contributor.utdAuthorBhat, Vandita D.
dc.contributor.utdAuthorShukla, Tarjani
dc.contributor.utdAuthorCampbell, Zachary T.
dc.date.accessioned2020-09-18T15:52:33Z
dc.date.available2020-09-18T15:52:33Z
dc.date.issued2019-01-17
dc.description.abstractPUF (PUmilio/FBF) RNA-binding proteins recognize distinct elements. In C. elegans, PUF-8 binds to an 8-nt motif and restricts proliferation in the germline. Conversely, FBF-2 recognizes a 9-nt element and promotes mitosis. To understand how motif divergence relates to biological function, we first determined a crystal structure of PUF-8. Comparison of this structure to that of FBF-2 revealed a major difference in a central repeat. We devised a modified yeast 3-hybrid screen to identify mutations that confer recognition of an 8-nt element to FBF-2. We identified several such mutants and validated structurally and biochemically their binding to 8-nt RNA elements. Using genome engineering, we generated a mutant animal with a substitution in FBF-2 that confers preferential binding to the PUF-8 element. The mutant largely rescued overproliferation in animals that spontaneously generate tumors in the absence of puf-8. This work highlights the critical role of motif length in the specification of biological function.
dc.description.departmentSchool of Natural Sciences and Mathematics
dc.description.sponsorshipNIH grants R01NS100788, RGM122001A; US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. W-31– 109-Eng-38.
dc.identifier.bibliographicCitationBhat, Vandita D., Kathleen L. McCann, Yeming Wang, Dallas R. Fonseca, et al. 2019. "Engineering a conserved RNA regulatory protein repurposes its biological function in vivo." eLife 8: art. e43788, doi: 10.7554/eLife.43788
dc.identifier.issn2050-084X
dc.identifier.urihttps://dx.doi.org/10.7554/eLife.43788
dc.identifier.urihttps://hdl.handle.net/10735.1/8906
dc.identifier.volume8
dc.language.isoen
dc.publishereLife Sciences Publications Ltd
dc.rightsCC0 1.0 (Public Domain Dedication)
dc.rightsNo Copyright
dc.rights.urihttps://creativecommons.org/publicdomain/zero/1.0/
dc.source.journaleLife
dc.subjectMitosis
dc.subjectLife sciences
dc.subject.meshPUF-8 protein, C elegans
dc.subject.meshfem-3-binding protein, C elegans
dc.subject.meshBiological Science Disciplines
dc.titleEngineering a Conserved RNA Regulatory Protein Repurposes its Biological Function in Vivo
dc.type.genrearticle

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