Characterization of a novel, testis-specific equine serine/threonine kinase

Khalida Sabeur, Barry A. Ball, C. Jo Corbin, Alan J Conley

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Testis-specific protein kinases are important because of their potential role in spermiogenesis, sperm maturation, and sperm function. In the present study, a novel serine-threonine kinase with high identity to human serine-threonine kinase 31 (STK31) was cloned from equine testis and expression of the protein was characterized in equine testis and ejaculated spermatozoa. Five over-lapping independent clones were plaque purified after screening of a λ ZAP cDNA expression library constructed from equine testis. Sequence analysis and alignment of all five clones showed high identity with human STK31 with approximately 200 bp of the equine N-terminal sequence incomplete. The putative full-length coding sequence of this testis specific equine cDNA was completed by amplification of a 200-bp fragment using a human primer upstream of the reported translational start site with equine specific nested primers. Northern blot analysis using the equine STK31 cDNA detected an RNA transcript of ∼3.1 kb present in testis but not in other reproductive or somatic tissues. Immunolocalization of the protein in equine testis and spermatozoa demonstrated that STK31 was present in post-meiotic germ cells with localization to the equatorial segment of testicular spermatozoa. Analysis of the domain structure of equine STK31 revealed a protein kinase domain along with a putative RNA-binding region. The post-meiotic expression of this protein along with its domain structure suggests that STK31 may have a role in reorganization of sperm chromatin during spermiogenesis. The cloning of this novel, testis-specific equine STK provides a new tool to explore the role of kinases in sperm function.

Original languageEnglish (US)
Pages (from-to)867-873
Number of pages7
JournalMolecular Reproduction and Development
Volume75
Issue number5
DOIs
StatePublished - May 2008

Fingerprint

Protein-Serine-Threonine Kinases
Horses
Testis
Spermatozoa
Spermatogenesis
Protein Kinases
Complementary DNA
Clone Cells
RNA
Sperm Maturation
Proteins
Sequence Alignment
Gene Library
Germ Cells
Northern Blotting
Chromatin
Sequence Analysis
Organism Cloning
Phosphotransferases

Keywords

  • Horse
  • Serine-threonine kinase
  • Spermatozoa
  • Testis

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Characterization of a novel, testis-specific equine serine/threonine kinase. / Sabeur, Khalida; Ball, Barry A.; Corbin, C. Jo; Conley, Alan J.

In: Molecular Reproduction and Development, Vol. 75, No. 5, 05.2008, p. 867-873.

Research output: Contribution to journalArticle

Sabeur, Khalida ; Ball, Barry A. ; Corbin, C. Jo ; Conley, Alan J. / Characterization of a novel, testis-specific equine serine/threonine kinase. In: Molecular Reproduction and Development. 2008 ; Vol. 75, No. 5. pp. 867-873.
@article{c4fb782e7a6345a492cdd649f107ee11,
title = "Characterization of a novel, testis-specific equine serine/threonine kinase",
abstract = "Testis-specific protein kinases are important because of their potential role in spermiogenesis, sperm maturation, and sperm function. In the present study, a novel serine-threonine kinase with high identity to human serine-threonine kinase 31 (STK31) was cloned from equine testis and expression of the protein was characterized in equine testis and ejaculated spermatozoa. Five over-lapping independent clones were plaque purified after screening of a λ ZAP cDNA expression library constructed from equine testis. Sequence analysis and alignment of all five clones showed high identity with human STK31 with approximately 200 bp of the equine N-terminal sequence incomplete. The putative full-length coding sequence of this testis specific equine cDNA was completed by amplification of a 200-bp fragment using a human primer upstream of the reported translational start site with equine specific nested primers. Northern blot analysis using the equine STK31 cDNA detected an RNA transcript of ∼3.1 kb present in testis but not in other reproductive or somatic tissues. Immunolocalization of the protein in equine testis and spermatozoa demonstrated that STK31 was present in post-meiotic germ cells with localization to the equatorial segment of testicular spermatozoa. Analysis of the domain structure of equine STK31 revealed a protein kinase domain along with a putative RNA-binding region. The post-meiotic expression of this protein along with its domain structure suggests that STK31 may have a role in reorganization of sperm chromatin during spermiogenesis. The cloning of this novel, testis-specific equine STK provides a new tool to explore the role of kinases in sperm function.",
keywords = "Horse, Serine-threonine kinase, Spermatozoa, Testis",
author = "Khalida Sabeur and Ball, {Barry A.} and Corbin, {C. Jo} and Conley, {Alan J}",
year = "2008",
month = "5",
doi = "10.1002/mrd.20792",
language = "English (US)",
volume = "75",
pages = "867--873",
journal = "Molecular Reproduction and Development",
issn = "1040-452X",
publisher = "Wiley-Liss Inc.",
number = "5",

}

TY - JOUR

T1 - Characterization of a novel, testis-specific equine serine/threonine kinase

AU - Sabeur, Khalida

AU - Ball, Barry A.

AU - Corbin, C. Jo

AU - Conley, Alan J

PY - 2008/5

Y1 - 2008/5

N2 - Testis-specific protein kinases are important because of their potential role in spermiogenesis, sperm maturation, and sperm function. In the present study, a novel serine-threonine kinase with high identity to human serine-threonine kinase 31 (STK31) was cloned from equine testis and expression of the protein was characterized in equine testis and ejaculated spermatozoa. Five over-lapping independent clones were plaque purified after screening of a λ ZAP cDNA expression library constructed from equine testis. Sequence analysis and alignment of all five clones showed high identity with human STK31 with approximately 200 bp of the equine N-terminal sequence incomplete. The putative full-length coding sequence of this testis specific equine cDNA was completed by amplification of a 200-bp fragment using a human primer upstream of the reported translational start site with equine specific nested primers. Northern blot analysis using the equine STK31 cDNA detected an RNA transcript of ∼3.1 kb present in testis but not in other reproductive or somatic tissues. Immunolocalization of the protein in equine testis and spermatozoa demonstrated that STK31 was present in post-meiotic germ cells with localization to the equatorial segment of testicular spermatozoa. Analysis of the domain structure of equine STK31 revealed a protein kinase domain along with a putative RNA-binding region. The post-meiotic expression of this protein along with its domain structure suggests that STK31 may have a role in reorganization of sperm chromatin during spermiogenesis. The cloning of this novel, testis-specific equine STK provides a new tool to explore the role of kinases in sperm function.

AB - Testis-specific protein kinases are important because of their potential role in spermiogenesis, sperm maturation, and sperm function. In the present study, a novel serine-threonine kinase with high identity to human serine-threonine kinase 31 (STK31) was cloned from equine testis and expression of the protein was characterized in equine testis and ejaculated spermatozoa. Five over-lapping independent clones were plaque purified after screening of a λ ZAP cDNA expression library constructed from equine testis. Sequence analysis and alignment of all five clones showed high identity with human STK31 with approximately 200 bp of the equine N-terminal sequence incomplete. The putative full-length coding sequence of this testis specific equine cDNA was completed by amplification of a 200-bp fragment using a human primer upstream of the reported translational start site with equine specific nested primers. Northern blot analysis using the equine STK31 cDNA detected an RNA transcript of ∼3.1 kb present in testis but not in other reproductive or somatic tissues. Immunolocalization of the protein in equine testis and spermatozoa demonstrated that STK31 was present in post-meiotic germ cells with localization to the equatorial segment of testicular spermatozoa. Analysis of the domain structure of equine STK31 revealed a protein kinase domain along with a putative RNA-binding region. The post-meiotic expression of this protein along with its domain structure suggests that STK31 may have a role in reorganization of sperm chromatin during spermiogenesis. The cloning of this novel, testis-specific equine STK provides a new tool to explore the role of kinases in sperm function.

KW - Horse

KW - Serine-threonine kinase

KW - Spermatozoa

KW - Testis

UR - http://www.scopus.com/inward/record.url?scp=41149107076&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41149107076&partnerID=8YFLogxK

U2 - 10.1002/mrd.20792

DO - 10.1002/mrd.20792

M3 - Article

C2 - 18246530

AN - SCOPUS:41149107076

VL - 75

SP - 867

EP - 873

JO - Molecular Reproduction and Development

JF - Molecular Reproduction and Development

SN - 1040-452X

IS - 5

ER -