Engineering a conserved RNA regulatory protein repurposes its biological function in vivo.
Elife
; 82019 01 17.
Article
en En
| MEDLINE
| ID: mdl-30652968
ABSTRACT
PUF (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.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Ingeniería de Proteínas
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Proteínas de Unión al ARN
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Caenorhabditis elegans
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Proteínas de Caenorhabditis elegans
Límite:
Animals
Idioma:
En
Año:
2019
Tipo del documento:
Article