Skip to main content

The Use of cDNA Libraries to Demonstrate a Linkage Between Transcription and Translation in Male Germ Cells

  • Chapter
  • 1030 Accesses

Abstract

cDNA libraries have played a prominent role in developing the extensive database of gene expression in germ cells and somatic cells of the mammalian testis. Differential screening of cDNA libraries has allowed investigators to determine the temporal up-and downregulation of many genes. This chapter discusses how suppressive subtraction hybridization and cDNA sequencing have been used to define populations of messenger RNAs (mRNAs) that selectively bind, or do not bind, to the germ cell-specific Y-box protein, MSY2. MSY2 is an abundant DNA/RNA-binding protein that in vitro binds to all mRNAs, but shows selective binding to a subset of male germ cell mRNAs in cells. This specificity is regulated by MSY2 binding to a conserved sequence in gene promoters, which facilitates MSY2 binding to the transcripts from these promoters in the nucleus and coordinates the transport, storage, and translational suppression of these mRNAs in the cytoplasm.

This is a preview of subscription content, log in via an institution.

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Eddy EM. Male germ cell gene expression. Recent Prog Horm Res 2002;57:103–128.

    Article  PubMed  CAS  Google Scholar 

  2. Shima JE, McLean DJ, McCarrey JR, Griswold MD. The murine testicular transcriptome: characterizing gene expression in the testis during the progression of spermatogenesis. Biol Reprod 2004;71:319–330.

    Article  PubMed  CAS  Google Scholar 

  3. Schlecht U, Demougin P, Koch R, et al. Expression profiling of mammalian male meiosis and gametogenesis identifies novel candidate genes for roles in the regulation of fertility. Mol Biol Cell 2004;15:1031–1043.

    Article  PubMed  CAS  Google Scholar 

  4. Wrobel G, Primig M. Mammalian male germ cells are fertile ground for expression profiling of sexual reproduction. Reproduction 2005;129:1–7.

    Article  PubMed  CAS  Google Scholar 

  5. Romrell LJ, Bellve AR, Fawcett DW. Separation of mouse spermatogenic cells by sedimentation velocity. A morphological characterization. Dev Biol 1976;49:119–131.

    Article  PubMed  CAS  Google Scholar 

  6. Grabske RJ, Lake S, Gledhill BL, Meistrich ML. Centrifugal elutriation: separation of spermatogenic cells on the basis of sedimentation velocity. J Cell Physiol 1975;86:177–189.

    Article  PubMed  CAS  Google Scholar 

  7. Monesi V, Geremia R, D’Agosino A, Boitani C. Biochemistry of male germ cell differentiation in mammals: RNA synthesis in meiotic and postmeiotic cells. Curr Top Dev Biol 1978;12:11–36.

    Article  PubMed  CAS  Google Scholar 

  8. Kleene KC, Distel RJ, Hecht NB. cDNA clones encoding poly (A) RNAs which first appear at detectable levels in haploid phases of spermatogenesis in the mouse. Dev Biol 1983;98:455–464.

    Article  PubMed  CAS  Google Scholar 

  9. Dudley K, Potter J, Lyon MF, Willison KR. Analysis of male sterile mutations in the mouse using haploid stage expressed cDNA probes. Nucleic Acids Res 1984;25:4281–4293.

    Article  Google Scholar 

  10. Fujii T, Tamura K, Masai K, Tanaka H, Nishimune Y, Nojima H. Use of stepwise subtraction to comprehensively isolate mouse genes whose transcription is upregulated during spermiogenesis. EMBO Rep 2002;3:367–372.

    Article  PubMed  CAS  Google Scholar 

  11. Hecht NB. Molecular mechanisms of male germ cell differentiation. BioEssays 1998;20:555–561.

    Article  PubMed  CAS  Google Scholar 

  12. Tanaka H, Baba T. Gene expression in spermiogenesis. Cell Mol Life Sci 2005;62:344–354.

    Article  PubMed  CAS  Google Scholar 

  13. Yang J, Medvedev S, Reddi PP, Schultz RM, Hecht NB. The DNA/RNA-binding protein MSY2 marks specific transcripts for cytoplasmic storage in mouse male germ cells. Proc Natl Acad Sci USA 2005;102:1513–1518.

    Article  PubMed  CAS  Google Scholar 

  14. Yiu GK, Hecht NB. Novel testis-specific protein-DNA interactions activate transcription of the mouse protamine 2 gene during spermatogenesis. J Biol Chem 1997;272:26,926–29,933.

    Article  PubMed  CAS  Google Scholar 

  15. Gu W, Tekur S, Reinbold R, et al. Mammalian male and female germ cells express a germ cell-specific Y-Box protein, MSY2. Biol Reprod 1998;59:1266–1274.

    Article  PubMed  CAS  Google Scholar 

  16. Yang Y, Medvedev S, Yu J, et al. Absence of the DNA/RNA-binding protein MSY2 results in male and female infertility. Proc Nat Acad Sci USA 2005;102:5755–5760.

    Article  PubMed  CAS  Google Scholar 

  17. Yu J, Hecht NB, Schultz RM. Expression of MSY2, a germ cell-specific Y-box protein in the female mouse. Biol Reprod 2001;65:1260–1270.

    Article  PubMed  CAS  Google Scholar 

  18. Yu J, Hecht NB, Schultz RM. RNA-binding properties and translation repression in vitro by germ cell-specific MSY2 protein. Biol Reprod 2002;67:1093–1098.

    Article  PubMed  CAS  Google Scholar 

  19. Tekur S, Pawlak A, Guellaen G, Hecht NB. Contrin, the human homologue of a germ-cell Y-box-binding protein: cloning, expression, and chromosomal localization. J Androl 1999;20:135–144.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this chapter

Cite this chapter

Hecht, N.B. (2007). The Use of cDNA Libraries to Demonstrate a Linkage Between Transcription and Translation in Male Germ Cells. In: Carrell, D.T. (eds) The Genetics of Male Infertility. Humana Press. https://doi.org/10.1007/978-1-59745-176-5_2

Download citation

Publish with us

Policies and ethics