Submitted March 18, 2008
Returned for revision March 24, 2008
Accepted April 1, 2008
Gamete Biology
Phosphorylation-Dependent Interaction of Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein (YWHA) with PADI6 Following Oocyte Maturation in Mice
Alan J. Snow ,
Pawan Puri ,
Amparo Acker-Palmer ,
Tewis Bouwmeester ,
Srinivasan Vijayaraghavan ,
and
Douglas Kline *
* To whom correspondence should be addressed. E-mail: dkline{at}kent.edu.
Abstract
Proteins in the tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein family (YWHA, also known as 14-3-3) are involved in the regulation of many intracellular processes. We have examined the interaction of YWHA with peptidylarginine deiminase, type VI (PADI6), an abundant protein found in mammalian oocytes, eggs and early embryos. Peptidylarginine deiminases catalyze the posttranslational modification of peptidylarginine to citrulline. PADI6 is associated with oocyte cytoplasmic sheets and it has been shown that PADI6-deficient mice are infertile due to disruption of development beyond the two-cell stage. We found that PADI6 undergoes a dramatic developmental change in phosphorylation during oocyte maturation. This change in phosphorylation is linked to an interaction of PADI6 with YWHA in the mature egg. Recombinant glutathione S-transferase (GST) YWHA pull-down experiments and transgenic tandem affinity purification with liquid chromatography-mass spectrometry demonstrate a binding interaction between YWHA and PADI6 in mature eggs. YWHA proteins modulate or complement intracellular events involving phosphorylation dependent switching or protein modification. These results indicate that phosphorylation and/or YWHA binding may serve as a means of intracellular PADI6 regulation.
Key words:
Oocyte development •
14-3-3 •
PADI6 •
oocyte maturation •
peptidylarginine deiminase
