A novel germ cell protein, SPIF (sperm PKA interacting factor), is essential for the formation of a PKA/TCP11 complex that undergoes conformational and phosphorylation changes upon capacitation

Simone J. Stanger, Estelle A. Law, Duangporn Jamsai, Moira K. O'Bryan, Brett Nixon, Eileen A. McLaughlin, R. John Aitken, Shaun D. Roman

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Spermatozoa require the process of capacitation to enable them to fertilize an egg. PKA is crucial to capacitation and the development of hyperactivated motility. Sperm PKA is activated by cAMP generated by the germ cell-enriched adenylyl cyclase encoded by Adcy10. Male mice lacking Adcy10 are sterile, because their spermatozoa are immotile. The current studywas designed to identify binding partners of the sperm-specific (Cα2) catalytic subunit of PKA (PRKACA) by using it as the "bait" in a yeast 2-hybrid system. This approach was used to identify a novel germ cell-enriched protein, sperm PKA interacting factor (SPIF), in 25% of the positive clones. Homozygous Spif-nullmice were embryonically lethal. SPIF was coexpressed and coregulated with PRKACA and with t-complex protein (TCP)-11, a protein associated with PKA signaling. We established that these 3 proteins form part of a novel complex in mouse spermatozoa. Upon capacitation, the SPIF protein becomes tyrosine phosphorylatedin > 95% of sperm. Anapparent molecular rearrangement in the complex occurs, bringing PRKACA and TCP11 into proximity. Taken together, these results suggest a role for the novel complex of SPIF, PRKACA, and TCP11 during spermcapacitation, fertilization, and embryogenesis.

Original languageEnglish
Pages (from-to)2777-2791
Number of pages15
JournalFASEB Journal
Volume30
Issue number8
DOIs
Publication statusPublished - 1 Aug 2016
Externally publishedYes

Fingerprint

Phosphorylation
Germ Cells
Spermatozoa
Cells
Proteins
Hybrid systems
Adenylyl Cyclases
Yeast
Tyrosine
Fertilization
Embryonic Development
Ovum
Catalytic Domain
Clone Cells
Yeasts

Cite this

Stanger, Simone J. ; Law, Estelle A. ; Jamsai, Duangporn ; O'Bryan, Moira K. ; Nixon, Brett ; McLaughlin, Eileen A. ; Aitken, R. John ; Roman, Shaun D. / A novel germ cell protein, SPIF (sperm PKA interacting factor), is essential for the formation of a PKA/TCP11 complex that undergoes conformational and phosphorylation changes upon capacitation. In: FASEB Journal. 2016 ; Vol. 30, No. 8. pp. 2777-2791.
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abstract = "Spermatozoa require the process of capacitation to enable them to fertilize an egg. PKA is crucial to capacitation and the development of hyperactivated motility. Sperm PKA is activated by cAMP generated by the germ cell-enriched adenylyl cyclase encoded by Adcy10. Male mice lacking Adcy10 are sterile, because their spermatozoa are immotile. The current studywas designed to identify binding partners of the sperm-specific (Cα2) catalytic subunit of PKA (PRKACA) by using it as the {"}bait{"} in a yeast 2-hybrid system. This approach was used to identify a novel germ cell-enriched protein, sperm PKA interacting factor (SPIF), in 25{\%} of the positive clones. Homozygous Spif-nullmice were embryonically lethal. SPIF was coexpressed and coregulated with PRKACA and with t-complex protein (TCP)-11, a protein associated with PKA signaling. We established that these 3 proteins form part of a novel complex in mouse spermatozoa. Upon capacitation, the SPIF protein becomes tyrosine phosphorylatedin > 95{\%} of sperm. Anapparent molecular rearrangement in the complex occurs, bringing PRKACA and TCP11 into proximity. Taken together, these results suggest a role for the novel complex of SPIF, PRKACA, and TCP11 during spermcapacitation, fertilization, and embryogenesis.",
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A novel germ cell protein, SPIF (sperm PKA interacting factor), is essential for the formation of a PKA/TCP11 complex that undergoes conformational and phosphorylation changes upon capacitation. / Stanger, Simone J.; Law, Estelle A.; Jamsai, Duangporn; O'Bryan, Moira K.; Nixon, Brett; McLaughlin, Eileen A.; Aitken, R. John; Roman, Shaun D.

In: FASEB Journal, Vol. 30, No. 8, 01.08.2016, p. 2777-2791.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A novel germ cell protein, SPIF (sperm PKA interacting factor), is essential for the formation of a PKA/TCP11 complex that undergoes conformational and phosphorylation changes upon capacitation

AU - Stanger, Simone J.

AU - Law, Estelle A.

AU - Jamsai, Duangporn

AU - O'Bryan, Moira K.

AU - Nixon, Brett

AU - McLaughlin, Eileen A.

AU - Aitken, R. John

AU - Roman, Shaun D.

PY - 2016/8/1

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AB - Spermatozoa require the process of capacitation to enable them to fertilize an egg. PKA is crucial to capacitation and the development of hyperactivated motility. Sperm PKA is activated by cAMP generated by the germ cell-enriched adenylyl cyclase encoded by Adcy10. Male mice lacking Adcy10 are sterile, because their spermatozoa are immotile. The current studywas designed to identify binding partners of the sperm-specific (Cα2) catalytic subunit of PKA (PRKACA) by using it as the "bait" in a yeast 2-hybrid system. This approach was used to identify a novel germ cell-enriched protein, sperm PKA interacting factor (SPIF), in 25% of the positive clones. Homozygous Spif-nullmice were embryonically lethal. SPIF was coexpressed and coregulated with PRKACA and with t-complex protein (TCP)-11, a protein associated with PKA signaling. We established that these 3 proteins form part of a novel complex in mouse spermatozoa. Upon capacitation, the SPIF protein becomes tyrosine phosphorylatedin > 95% of sperm. Anapparent molecular rearrangement in the complex occurs, bringing PRKACA and TCP11 into proximity. Taken together, these results suggest a role for the novel complex of SPIF, PRKACA, and TCP11 during spermcapacitation, fertilization, and embryogenesis.

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KW - Sperm capacitation

KW - Tyrosine kinase

KW - Spermatozoa/physiology

KW - Protein Subunits

KW - Phosphorylation

KW - Membrane Proteins/genetics

KW - Male

KW - Cyclic AMP-Dependent Protein Kinases/genetics

KW - RNA, Messenger/genetics

KW - Sex Ratio

KW - Sperm Capacitation/physiology

KW - Two-Hybrid System Techniques

KW - Animals

KW - Carrier Proteins/genetics

KW - Protein Isoforms

KW - Female

KW - Heterozygote

KW - Protein Conformation

KW - Mice

KW - Gene Expression Regulation/physiology

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U2 - 10.1096/fj.201500136R

DO - 10.1096/fj.201500136R

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