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  • CNOT6 regulates a novel pattern of mRNA deadenylation during oocyte meiotic maturation.

CNOT6 regulates a novel pattern of mRNA deadenylation during oocyte meiotic maturation.

Scientific reports (2018-05-03)
Karl-Frédéric Vieux, Hugh J Clarke
ABSTRACT

In many cell types, the length of the poly(A) tail of an mRNA is closely linked to its fate - a long tail is associated with active translation, a short tail with silencing and degradation. During mammalian oocyte development, two contrasting patterns of polyadenylation have been identified. Some mRNAs carry a long poly(A) tail during the growth stage and are actively translated, then become deadenylated and down-regulated during the subsequent stage, termed meiotic maturation. Other mRNAs carry a short tail poly(A) tail and are translationally repressed during growth, and their poly(A) tail lengthens and they become translationally activated during maturation. As well, a program of elimination of this 'maternal' mRNA is initiated during oocyte maturation. Here we describe a third pattern of polyadenylation: mRNAs are deadenylated in growing oocytes, become polyadenylated during early maturation and then deadenylated during late maturation. We show that the deadenylase, CNOT6, is present in cortical foci of oocytes and regulates deadenylation of these mRNAs, and that PUF-binding elements (PBEs) regulate deadenylation in mature oocytes. Unexpectedly, maintaining a long poly(A) tail neither enhances translation nor inhibits degradation of these mRNAs. Our findings implicate multiple machineries, more complex than previously thought, in regulating mRNA activity in oocytes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-CNOT7 antibody produced in mouse, clone 2F6, purified immunoglobulin, buffered aqueous solution
Grace Bio-Labs SecureSeal imaging spacer, 8 wells, diam. × thickness 9 mm × 0.12 mm
Sigma-Aldrich
Phalloidin–Tetramethylrhodamine B isothiocyanate, sequence from Amanita phalloides(synthetic: peptide sequence)
Sigma-Aldrich
Anti-α-Tubulin antibody, Mouse monoclonal, clone AA13, purified from hybridoma cell culture
Sigma-Aldrich
Adenosine 3′,5′-cyclic monophosphate tris salt, ≥97% (HPLC), powder