6NOD

Crystal structure of C. elegans PUF-8 in complex with RNA

Summary for 6NOD

• Entry DOI: 10.2210/pdb6nod/pdb
• Related: 6NOC
• Descriptor: PUF (Pumilio/FBF) domain-containing, RNA (5'-R(P*UP*GP*UP*AP*UP*AP*UP*A)-3') (3 entities in total)
• Functional Keywords: pum repeat protein, rna binding protein, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Caenorhabditis elegans; More
• Total number of polymer chains: 6
• Total formula weight: 131210.24

Authors

Wang, Y.,McCann, K.L.,Qiu, C.,Hall, T.M.T. (deposition date: 2019-01-16, release date: 2019-01-30, Last modification date: 2024-03-13)

Primary citation

Bhat, V.D.,McCann, K.L.,Wang, Y.,Fonseca, D.R.,Shukla, T.,Alexander, J.C.,Qiu, C.,Wickens, M.,Lo, T.W.,Tanaka Hall, T.M.,Campbell, Z.T.
Engineering a conserved RNA regulatory protein repurposes its biological function in vivo .
Elife, 8:-, 2019

PubMed Abstract: PUF (milio/BF) RNA-binding proteins recognize distinct elements. In , 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 . This work highlights the critical role of motif length in the specification of biological function.

PubMed: 30652968
DOI: 10.7554/eLife.43788

Experimental method

X-RAY DIFFRACTION (2.547 Å)