Skip to main content

Research Repository

Advanced Search

The enzyme activities of Caf1 and Ccr4 are both required for deadenylation by the human Ccr4-Not nuclease module

Maryati, Marayti; Airhihen, Blessing; Winkler, G. Sebastiaan

Authors

Marayti Maryati

Blessing Airhihen



Abstract

In eukaryotic cells, the shortening and removal of the poly(A) tail (deadenylation) of cytoplasmic mRNA is a key event in regulated mRNA degradation. A major enzyme involved in deadenylation is the Ccr4-Not deadenylase complex, which can be recruited to its target mRNA by RNA-binding proteins or the miRNA repression complex. In addition to six non-catalytic components, the complex contains two enzymatic subunits with ribonuclease activity: Ccr4 and Caf1 (Pop2). In vertebrates, each deadenylase subunit is encoded by two paralogues: Caf1, which can interact with the anti-proliferative protein BTG2, is encoded by CNOT7 and CNOT8, while Ccr4 is encoded by the highly similar genes CNOT6 and CNOT6L. Currently, it is unclear whether the catalytic subunits work cooperatively, or whether the nuclease components have unique roles in deadenylation. We therefore developed a method to express and purify a minimal human BTG2-Caf1-Ccr4 nuclease sub-complex from bacterial cells. By using chemical inhibition and well-characterised inactivating amino acid substitutions, we demonstrate that the enzyme activities of Caf1 and Ccr4 are both required for deadenylation in vitro. These results indicate that Caf1 and Ccr4 cooperate in mRNA deadenylation and suggest that the enzyme activities of Caf1 and Ccr4 are regulated via allosteric interactions within the nuclease module.

Journal Article Type Article
Publication Date Jun 19, 2015
Journal Biochemical Journal
Print ISSN 0264-6021
Electronic ISSN 0264-6021
Publisher Portland Press
Peer Reviewed Peer Reviewed
Volume 469
Issue 1
APA6 Citation Maryati, M., Airhihen, B., & Winkler, G. S. (2015). The enzyme activities of Caf1 and Ccr4 are both required for deadenylation by the human Ccr4-Not nuclease module. Biochemical Journal, 469(1), doi:10.1042/BJ20150304
DOI https://doi.org/10.1042/BJ20150304
Keywords Ccr4-Not; ribonuclease; poly(A); deadenylase; mRNA decay; post-transcriptional gene regulation
Publisher URL http://www.biochemj.org/content/469/1/169
Copyright Statement Copyright information regarding this work can be found at the following address: http://eprints.nottingh.../end_user_agreement.pdf
Additional Information This research was originally published in Biochemical Journal. Maryati Maryati, Blessing Airhihen, G. Sebastiaan Winkler, The enzyme activities of Caf1 and Ccr4 are both required for deadenylation by the human Ccr4–Not nuclease module. Biochemical Journal. 2015; v. 469: pp169-176 © the Biochemical Society

Files

bj20150304 Portland press revised.pdf (753 Kb)
PDF

Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf





You might also like



Downloadable Citations

;