Brocard, Michele, Khasnis, Sarika Jayant, Wood, C David, Shannon-Lowe, Claire and West, Michelle J (2018) Pumilio directs deadenylation-associated translational repression of the cyclin-dependent kinase 1 activator RGC-32. Nucleic Acids Research, 46 (7). pp. 3707-3725. ISSN 0305-1048

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Abstract

Response gene to complement-32 (RGC-32) activates cyclin-dependent kinase 1, regulates the cell cycle and is deregulated in many human tumours. We previously showed that RGC-32 expression is upregulated by the cancer-associated Epstein-Barr virus (EBV) in latently infected B cells through the relief of translational repression. We now show that EBV infection of naïve primary B cells also induces RGC-32 protein translation. In EBV-immortalised cell lines, we found that RGC-32 depletion resulted in cell death, indicating a key role in B cell survival. Studying RGC-32 translational control in EBV-infected cells, we found that the RGC-32 3′untranslated region (3′UTR) mediates translational repression. Repression was dependent on a single Pumilio binding element (PBE) adjacent to the polyadenylation signal. Mutation of this PBE did not affect mRNA cleavage, but resulted in increased polyA tail length. Consistent with Pumilio-dependent recruitment of deadenylases, we found that depletion of Pumilio in EBV-infected cells increased RGC-32 protein expression and polyA tail length. The extent of Pumilio binding to the endogenous RGC-32 mRNA in EBV-infected cell lines also correlated with RGC-32 protein expression. Our data demonstrate the importance of RGC-32 for the survival of EBV-immortalised B cells and identify Pumilio as a key regulator of RGC-32 translation.

Item Type:	Article
Keywords:	virus, cell cycle, translation, deadenylation, Pumilio
Schools and Departments:	School of Life Sciences > Biochemistry
Research Centres and Groups:	Gene Expression Research Group Haematology Research Group
Subjects:	Q Science > QH Natural history > QH0301 Biology > QH0573 Cytology > QH0605 Cell division Q Science > QP Physiology > QP0501 Animal biochemistry > QP0551 Proteins, amino acids, etc. Q Science > QP Physiology > QP0501 Animal biochemistry > QP0620 Nucleic acids Q Science > QR Microbiology Q Science > QR Microbiology > QR0355 Virology
Depositing User:	Michelle West
Date Deposited:	30 Jan 2018 09:03
Last Modified:	01 Jul 2019 18:31
URI:	http://sro.sussex.ac.uk/id/eprint/73206

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