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 lambda 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 approximately 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.
(c) 2008 Wiley-Liss, Inc.
Publication Date: 2008-02-05 PubMed ID: 18246530DOI: 10.1002/mrd.20792Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article discusses the identification and study of a new serine-threonine kinase identified in horse testes, which may play a crucial role in sperm maturation and function.
Introduction and Objective
- The focus was on protein kinases specific to the testes, which are considered to significantly impact spermiogenesis, sperm maturation, and sperm function.
- The study aimed at cloning a novel serine/threonine kinase from equine testis that showed high similarity to human serine-threonine kinase 31 (STK31).
- The researchers also aimed to characterize the expression of this protein in equine testis and ejaculated spermatozoa.
Methods Employed
- A lambda ZAP cDNA expression library built from equine testis was screened, resulting in five overlapping independent clones being plaque purified.
- Through sequence analysis and alignment of all these clones, high similarity was identified with human STK31.
- The researchers also amplified a 200-bp fragment using a human primer to complete the supposed full-length coding sequence of this testis-specific equine cDNA.
- Using the equine STK31 cDNA, a Northern blot analysis was executed which helped detect an RNA transcript only present in the testis but not in other reproductive or somatic tissues.
Findings from the Research
- The immunolocalization of the protein in equine testis and spermatozoa revealed that STK31 is found in post-meiotic germ cells.
- This protein localizes to the equatorial segment of testicular spermatozoa.
- The investigation into the domain structure of equine STK31 unveiled a protein kinase domain and a putative RNA-binding region.
Conclusion of the Study
- The presence of this protein in post-meiotic cells and its domain structure suggests a possible role of STK31 in reorganizing sperm chromatin during spermiogenesis.
- The cloning of this novel, testis-specific equine STK provides a vital tool for further exploring the role of kinases in sperm function.
Cite This Article
APA
Sabeur K, Ball BA, Corbin CJ, Conley A.
(2008).
Characterization of a novel, testis-specific equine serine/threonine kinase.
Mol Reprod Dev, 75(5), 867-873.
https://doi.org/10.1002/mrd.20792 Publication
Researcher Affiliations
- Department of Population Health and Reproduction, University of California Davis, Davis, California 95616, USA.
MeSH Terms
- Animals
- Chromatin Assembly and Disassembly / physiology
- Cloning, Molecular
- Gene Library
- Horses
- Humans
- Male
- Open Reading Frames / genetics
- Organ Specificity / physiology
- Protein Serine-Threonine Kinases / biosynthesis
- Protein Serine-Threonine Kinases / genetics
- Protein Structure, Tertiary / physiology
- RNA-Binding Proteins / biosynthesis
- RNA-Binding Proteins / genetics
- Sequence Homology, Nucleic Acid
- Spermatogenesis / physiology
- Spermatozoa / cytology
- Spermatozoa / enzymology
- Testis / cytology
- Testis / enzymology
Citations
This article has been cited 10 times.- Bae DH, Kim HJ, Yoon BH, Park JL, Kim M, Kim SK, Kim SY, Lee SI, Song KS, Kim YS. STK31 upregulation is associated with chromatin remodeling in gastric cancer and induction of tumorigenicity in a xenograft mouse model. Oncol Rep 2021 Apr;45(4).
- Luo S, Gao X, Ding J, Liu C, Du C, Hou C, Zhu J, Lou B. Transcriptome Sequencing Reveals the Traits of Spermatogenesis and Testicular Development in Large Yellow Croaker (Larimichthys crocea). Genes (Basel) 2019 Nov 21;10(12).
- Zhong L, Liu J, Hu Y, Wang W, Xu F, Xu W, Han J, Biskup E. STK31 as novel biomarker of metastatic potential and tumorigenicity of colorectal cancer. Oncotarget 2017 Apr 11;8(15):24354-24361.
- Jeong J, Jin S, Choi H, Kwon JT, Kim J, Kim J, Park ZY, Cho C. Characterization of MAGEG2 with testis-specific expression in mice. Asian J Androl 2017 Nov-Dec;19(6):659-665.
- Wu C, Crowhurst RN, Dennis AB, Twort VG, Liu S, Newcomb RD, Ross HA, Buckley TR. De Novo Transcriptome Analysis of the Common New Zealand Stick Insect Clitarchus hookeri (Phasmatodea) Reveals Genes Involved in Olfaction, Digestion and Sexual Reproduction. PLoS One 2016;11(6):e0157783.
- Stefaniuk M, Ropka-Molik K. RNA sequencing as a powerful tool in searching for genes influencing health and performance traits of horses. J Appl Genet 2016 May;57(2):199-206.
- Zhou J, Leu NA, Eckardt S, McLaughlin KJ, Wang PJ. STK31/TDRD8, a germ cell-specific factor, is dispensable for reproduction in mice. PLoS One 2014;9(2):e89471.
- Bao J, Wang L, Lei J, Hu Y, Liu Y, Shen H, Yan W, Xu C. STK31(TDRD8) is dynamically regulated throughout mouse spermatogenesis and interacts with MIWI protein. Histochem Cell Biol 2012 Mar;137(3):377-89.
- Sonenshine DE, Bissinger BW, Egekwu N, Donohue KV, Khalil SM, Roe RM. First transcriptome of the testis-vas deferens-male accessory gland and proteome of the spermatophore from Dermacentor variabilis (Acari: Ixodidae). PLoS One 2011;6(9):e24711.
- Wei Z, Wang Y, Zheng K, Wang Z, Liu R, Wang P, Li Y, Gao P, Akbari OS, Yang X. Loss-of-function in testis-specific serine/threonine protein kinase triggers male infertility in an invasive moth. Commun Biol 2024 Oct 4;7(1):1256.
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