The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity

Petar N. Grozdanov, Elahe Masoumzadeh, Michael P. Latham, Clinton C. MacDonald

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Cleavage and polyadenylation (C/P) of mRNA is an important cellular process that promotes increased diversity of mRNA isoforms and could change their stability in different cell types. The cleavage stimulation factor (CstF) complex, part of the C/P machinery, binds to U- and GU-rich sequences located downstream from the cleavage site through its RNAbinding subunit, CstF-64. Less is known about the function of the other two subunits of CstF, CstF- 77 and CstF-50. Here, we show that the carboxyterminus of CstF-77 plays a previously unrecognized role in enhancing C/P by altering how the RNA recognition motif (RRM) of CstF-64 binds RNA. In support of this finding, we also show that CstF-64 relies on CstF-77 to be transported to the nucleus; excess CstF-64 localizes to the cytoplasm, possibly via interaction with cytoplasmic RNAs. Reverse genetics and nuclear magnetic resonance studies of recombinant CstF-64 (RRM-Hinge) and CstF-77 (monkeytailcarboxy- terminal domain) indicate that the last 30 amino acids of CstF-77 increases the stability of the RRM, thus altering the affinity of the complex for RNA. These results provide new insights into the mechanism by which CstF regulates the location of the RNA cleavage site during C/P.

Original languageEnglish
Pages (from-to)12022-12039
Number of pages18
JournalNucleic Acids Research
Volume46
Issue number22
DOIs
StatePublished - 2018

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