RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility

Moira K. O'Bryan, Brett J. Clark, Eileen A. McLaughlin, Rebecca J. D'Sylva, Liza O'Donnell, Jacqueline A. Wilce, Jessie Sutherland, Anne E. O'Connor, Belinda Whittle, Christopher C. Goodnow, Christopher J. Ormandy, Duangporn Jamsai

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Alternative splicing of precursor messenger RNA (pre-mRNA) is common in mammalian cells and enables the production of multiple gene products from a single gene, thus increasing transcriptome and proteome diversity. Disturbance of splicing regulation is associated with many human diseases; however, key splicing factors that control tissue-specific alternative splicing remain largely undefined. In an unbiased genetic screen for essential male fertility genes in the mouse, we identified the RNA binding protein RBM5 (RNA binding motif 5) as an essential regulator of haploid male germ cell pre-mRNA splicing and fertility. Mice carrying a missense mutation (R263P) in the second RNA recognition motif (RRM) of RBM5 exhibited spermatid differentiation arrest, germ cell sloughing and apoptosis, which ultimately led to azoospermia (no sperm in the ejaculate) and male sterility. Molecular modelling suggested that the R263P mutation resulted in compromised mRNA binding. Within the adult mouse testis, RBM5 localises to somatic and germ cells including spermatogonia, spermatocytes and round spermatids. Through the use of RNA pull down coupled with microarrays, we identified 11 round spermatid-expressed mRNAs as putative RBM5 targets. Importantly, the R263P mutation affected pre-mRNA splicing and resulted in a shift in the isoform ratios, or the production of novel spliced transcripts, of most targets. Microarray analysis of isolated round spermatids suggests that altered splicing of RBM5 target pre-mRNAs affected expression of genes in several pathways, including those implicated in germ cell adhesion, spermatid head shaping, and acrosome and tail formation. In summary, our findings reveal a critical role for RBM5 as a pre-mRNA splicing regulator in round spermatids and male fertility. Our findings also suggest that the second RRM of RBM5 is pivotal for appropriate pre-mRNA splicing.

Original languageEnglish
Article number1003628
Pages (from-to)1-13
Number of pages13
JournalPLoS Genetics
Volume9
Issue number7
DOIs
Publication statusPublished - Jul 2013
Externally publishedYes

Fingerprint

germ cell
Spermatids
male fertility
spermatids
Germ Cells
Fertility
RNA
fertility
germ cells
RNA splicing
RNA Splicing
RNA Precursors
Alternative Splicing
alternative splicing
Genes
mice
Acrosome
Azoospermia
Messenger RNA
Spermatogonia

Cite this

O'Bryan, M. K., Clark, B. J., McLaughlin, E. A., D'Sylva, R. J., O'Donnell, L., Wilce, J. A., ... Jamsai, D. (2013). RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility. PLoS Genetics, 9(7), 1-13. [1003628]. https://doi.org/10.1371/journal.pgen.1003628
O'Bryan, Moira K. ; Clark, Brett J. ; McLaughlin, Eileen A. ; D'Sylva, Rebecca J. ; O'Donnell, Liza ; Wilce, Jacqueline A. ; Sutherland, Jessie ; O'Connor, Anne E. ; Whittle, Belinda ; Goodnow, Christopher C. ; Ormandy, Christopher J. ; Jamsai, Duangporn. / RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility. In: PLoS Genetics. 2013 ; Vol. 9, No. 7. pp. 1-13.
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O'Bryan, MK, Clark, BJ, McLaughlin, EA, D'Sylva, RJ, O'Donnell, L, Wilce, JA, Sutherland, J, O'Connor, AE, Whittle, B, Goodnow, CC, Ormandy, CJ & Jamsai, D 2013, 'RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility', PLoS Genetics, vol. 9, no. 7, 1003628, pp. 1-13. https://doi.org/10.1371/journal.pgen.1003628

RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility. / O'Bryan, Moira K.; Clark, Brett J.; McLaughlin, Eileen A.; D'Sylva, Rebecca J.; O'Donnell, Liza; Wilce, Jacqueline A.; Sutherland, Jessie; O'Connor, Anne E.; Whittle, Belinda; Goodnow, Christopher C.; Ormandy, Christopher J.; Jamsai, Duangporn.

In: PLoS Genetics, Vol. 9, No. 7, 1003628, 07.2013, p. 1-13.

Research output: Contribution to journalArticle

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O'Bryan MK, Clark BJ, McLaughlin EA, D'Sylva RJ, O'Donnell L, Wilce JA et al. RBM5 Is a Male Germ Cell Splicing Factor and Is Required for Spermatid Differentiation and Male Fertility. PLoS Genetics. 2013 Jul;9(7):1-13. 1003628. https://doi.org/10.1371/journal.pgen.1003628