PAWP, a sperm-specific WW domain-binding protein, promotes meiotic resumption and pronuclear development during fertilization

Alexander T H Wu, Peter Sutovsky, Gaurishankar Manandhar, Wei Xu, Mika Katayama, Billy N. Day, Kwang Wook Park, Young Joo Yi, Wei Xi Yan, Randall S. Prather, Richard Oko

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

We report a novel alkaline extractable protein of the sperm head that exclusively resides in the post-acrosomal sheath region of the perinuclear theca (PT) and is expressed and assembled in elongating spermatids. It is a protein that shares sequence homology to the N-terminal half of WW domain-binding protein 2, while the C-terminal half is unique and rich in proline. A functional PPXY consensus binding site for group-I WW domain-containing proteins, and numerous unique repeating motifs, YGXPPXG, are identified in the proline-rich region. Considering these molecular characteristics, we designated this protein PAWP for postacrosomal sheath WW domain-binding protein. Microinjection of recombinant PAWP or alkaline PT extract into metaphase II-arrested porcine, bovine, macaque, and Xenopus oocytes induced a high rate of pronuclear formation, which was prevented by co-injection of a competitive PPXY motif containing peptide derived from PAWP but not by co-injection of the point-mutated peptide. Intracytoplasmic sperm injection (ICSI) of porcine oocytes combined with co-injection of the competitive PPXY peptide or an anti-recombinant PAWP antiserum prevented pronuclear formation and arrested fertilization. Conversely, co-injection of the modified PPXY peptide, when the tyrosine residue of PPXY was either phosphorylated or substituted with phenylalanine, did not prevent ICSI-induced fertilization. This study uncovers a group IWW domain module signal transduction event within the fertilized egg that appears compulsory for meiotic resumption and pronuclear development during egg activation and provides compelling evidence that a PPXY motif of sperm-contributed PAWP can trigger these events.

Original languageEnglish
Pages (from-to)12164-12175
Number of pages12
JournalJournal of Biological Chemistry
Volume282
Issue number16
DOIs
Publication statusPublished - Apr 20 2007
Externally publishedYes

Fingerprint

Fertilization
Spermatozoa
Carrier Proteins
Peptides
Injections
Intracytoplasmic Sperm Injections
Proline
Oocytes
Proteins
Swine
Sperm Head
Amino Acid Sequence Homology
Signal transduction
Spermatids
Zygote
Microinjections
Macaca
Metaphase
Xenopus
Phenylalanine

ASJC Scopus subject areas

  • Biochemistry

Cite this

PAWP, a sperm-specific WW domain-binding protein, promotes meiotic resumption and pronuclear development during fertilization. / Wu, Alexander T H; Sutovsky, Peter; Manandhar, Gaurishankar; Xu, Wei; Katayama, Mika; Day, Billy N.; Park, Kwang Wook; Yi, Young Joo; Yan, Wei Xi; Prather, Randall S.; Oko, Richard.

In: Journal of Biological Chemistry, Vol. 282, No. 16, 20.04.2007, p. 12164-12175.

Research output: Contribution to journalArticle

Wu, ATH, Sutovsky, P, Manandhar, G, Xu, W, Katayama, M, Day, BN, Park, KW, Yi, YJ, Yan, WX, Prather, RS & Oko, R 2007, 'PAWP, a sperm-specific WW domain-binding protein, promotes meiotic resumption and pronuclear development during fertilization', Journal of Biological Chemistry, vol. 282, no. 16, pp. 12164-12175. https://doi.org/10.1074/jbc.M609132200
Wu, Alexander T H ; Sutovsky, Peter ; Manandhar, Gaurishankar ; Xu, Wei ; Katayama, Mika ; Day, Billy N. ; Park, Kwang Wook ; Yi, Young Joo ; Yan, Wei Xi ; Prather, Randall S. ; Oko, Richard. / PAWP, a sperm-specific WW domain-binding protein, promotes meiotic resumption and pronuclear development during fertilization. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 16. pp. 12164-12175.
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AU - Sutovsky, Peter

AU - Manandhar, Gaurishankar

AU - Xu, Wei

AU - Katayama, Mika

AU - Day, Billy N.

AU - Park, Kwang Wook

AU - Yi, Young Joo

AU - Yan, Wei Xi

AU - Prather, Randall S.

AU - Oko, Richard

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N2 - We report a novel alkaline extractable protein of the sperm head that exclusively resides in the post-acrosomal sheath region of the perinuclear theca (PT) and is expressed and assembled in elongating spermatids. It is a protein that shares sequence homology to the N-terminal half of WW domain-binding protein 2, while the C-terminal half is unique and rich in proline. A functional PPXY consensus binding site for group-I WW domain-containing proteins, and numerous unique repeating motifs, YGXPPXG, are identified in the proline-rich region. Considering these molecular characteristics, we designated this protein PAWP for postacrosomal sheath WW domain-binding protein. Microinjection of recombinant PAWP or alkaline PT extract into metaphase II-arrested porcine, bovine, macaque, and Xenopus oocytes induced a high rate of pronuclear formation, which was prevented by co-injection of a competitive PPXY motif containing peptide derived from PAWP but not by co-injection of the point-mutated peptide. Intracytoplasmic sperm injection (ICSI) of porcine oocytes combined with co-injection of the competitive PPXY peptide or an anti-recombinant PAWP antiserum prevented pronuclear formation and arrested fertilization. Conversely, co-injection of the modified PPXY peptide, when the tyrosine residue of PPXY was either phosphorylated or substituted with phenylalanine, did not prevent ICSI-induced fertilization. This study uncovers a group IWW domain module signal transduction event within the fertilized egg that appears compulsory for meiotic resumption and pronuclear development during egg activation and provides compelling evidence that a PPXY motif of sperm-contributed PAWP can trigger these events.

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