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BOR - Papers in Press, published online ahead of print November 21, 2007.
Biol Reprod 2007, 10.1095/biolreprod.107.064774
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BIOLOGY OF REPRODUCTION 78, 472–482 (2008)
DOI: 10.1095/biolreprod.107.064774
© 2008 by the Society for the Study of Reproduction, Inc.

Pre-Messenger RNA Cleavage Factor I (CFIm): Potential Role in Alternative Polyadenylation During Spermatogenesis1

Becky L. Sartini 3 5, Hang Wang 4 5, Wei Wang 5, Clarke F. Millette 6, and Daniel L. Kilpatrick 2 5

Department of Molecular and Cellular Physiology,5 University of Massachusetts Medical School, Worcester, Massachusetts 01655 Department of Cell and Developmental Biology and Anatomy,6 University of South Carolina School of Medicine, Columbia, South Carolina 29208

ABSTRACT

A hallmark of male germ cell gene expression is the generation by alternative polyadenylation of cell-specific mRNAs, many of which utilize noncanonical A(A/U)UAAA-independent polyadenylation signals. Cleavage factor I (CFIm), a component of the pre-mRNA cleavage and polyadenylation protein complex, can direct A(A/U)UAAA-independent polyadenylation site selection of somatic cell mRNAs. Here we report that the CFIm subunits NUDT21/CPSF5 and CPSF6 are highly enriched in mouse male germ cells relative to somatic cells. Both subunits are expressed from spermatogenic cell mRNAs that are shorter than the corresponding somatic transcripts. Complementary DNA sequencing and Northern blotting revealed that the shorter Nudt21 and Cpsf6 mRNAs are generated by alternative polyadenylation in male germ cells using proximal poly(A) signals. Both sets of transcripts contain CFIm binding sites within their 3'-untranslated regions, suggesting autoregulation of CFIm subunit formation in male germ cells. CFIm subunit mRNA and protein levels exhibit distinct developmental variation during spermatogenesis, indicating stage-dependent translational and/or posttranslational regulation. CFIm binding sites were identified near the 3' ends of numerous male germ cell transcripts utilizing A(A/U)UAAA-independent sites. Together these findings suggest that CFIm complexes participate in alternative polyadenylation directed by noncanonical poly(A) signals during spermatogenesis.

alternative polyadenylation, CFIm, gamete biology, gene regulation, spermatogenesis, testis

FOOTNOTES

3Current address: Department of Fisheries, Animal and Veterinary Science, University of Rhode Island, Kingston, RI 02881.

4Current address: Department of Urology, The Johns Hopkins University School of Medicine, Baltimore, MD 21287.

1Supported by Public Service grant R01 HD45723. Core resources supported by the Diabetes Endocrinology Research Center grant DK32520 were also used.

Correspondence: 2FAX: 508 856 5997; e-mail: daniel.kilpatrick{at}umassmed.edu






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Copyright © 2008 by the Society for the Study of Reproduction.