Cleavage of rRNA ensures translational cessation in sperm at fertilization

G. D. Johnson, E. Sendler, C. Lalancette, R. Hauser, M. P. Diamond, S. A. Krawetz

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Intact ribosomal RNAs (rRNAs) comprise the majority of somatic transcripts, yet appear conspicuously absent in spermatozoa, perhaps reflecting cytoplasmic expulsion during spermatogenesis. To discern their fate, total RNA retained in mature spermatozoa from three fertile donors was characterized by Next Generation Sequencing. In all samples, >75% of total sequence reads aligned to rRNAs. The distribution of reads along the length of these transcripts exhibited a high degree of non-uniformity that was reiterated between donors. The coverage of sequencing reads was inversely correlated with guanine-cytosine (GC)-richness such that sequences greater than ~70% GC were virtually absent in all sperm RNA samples. To confirm the loss of sequence, the relative abundance of specific regions of the 28S transcripts in sperm was established by 7-Deaza-2′-deoxy-guanosine-5′-triphosphate RT-PCR. The inability to amplify specific regions of the 28S sequence from sperm despite the abundant representation of this transcript in the sequencing libraries demonstrates that approximately three-quarters of RNA retained in the mature male gamete are products of rRNA fragmentation. Hence, cleavage (not expulsion of the RNA component of the translational machinery) is responsible for preventing spurious translation following spermiogenesis. These results highlight the potential importance of those transcripts, including many mRNAs, which evade fragmentation and remain intact when sperm are delivered at fertilization.

Original languageEnglish (US)
Article numbergar054
Pages (from-to)721-726
Number of pages6
JournalMolecular Human Reproduction
Volume17
Issue number12
DOIs
StatePublished - Dec 1 2011

Fingerprint

Ribosomal RNA
Fertilization
Spermatozoa
Cytosine
Guanine
Spermatogenesis
Deoxyguanosine
Guanosine Triphosphate
Germ Cells
Polymerase Chain Reaction
Messenger RNA

Keywords

  • Next generation sequencing
  • Sperm
  • Spermiogenesis
  • rRNA

ASJC Scopus subject areas

  • Reproductive Medicine
  • Embryology
  • Molecular Biology
  • Genetics
  • Obstetrics and Gynecology
  • Developmental Biology
  • Cell Biology

Cite this

Johnson, G. D., Sendler, E., Lalancette, C., Hauser, R., Diamond, M. P., & Krawetz, S. A. (2011). Cleavage of rRNA ensures translational cessation in sperm at fertilization. Molecular Human Reproduction, 17(12), 721-726. [gar054]. https://doi.org/10.1093/molehr/gar054

Cleavage of rRNA ensures translational cessation in sperm at fertilization. / Johnson, G. D.; Sendler, E.; Lalancette, C.; Hauser, R.; Diamond, M. P.; Krawetz, S. A.

In: Molecular Human Reproduction, Vol. 17, No. 12, gar054, 01.12.2011, p. 721-726.

Research output: Contribution to journalArticle

Johnson, GD, Sendler, E, Lalancette, C, Hauser, R, Diamond, MP & Krawetz, SA 2011, 'Cleavage of rRNA ensures translational cessation in sperm at fertilization', Molecular Human Reproduction, vol. 17, no. 12, gar054, pp. 721-726. https://doi.org/10.1093/molehr/gar054
Johnson, G. D. ; Sendler, E. ; Lalancette, C. ; Hauser, R. ; Diamond, M. P. ; Krawetz, S. A. / Cleavage of rRNA ensures translational cessation in sperm at fertilization. In: Molecular Human Reproduction. 2011 ; Vol. 17, No. 12. pp. 721-726.
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