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BOR - Papers in Press, published online ahead of print June 9, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.030130
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BIOLOGY OF REPRODUCTION 71, 1158–1166 (2004)
DOI: 10.1095/biolreprod.104.030130
© 2004 by the Society for the Study of Reproduction, Inc.

Pituitary

The 3' Untranslated Region of Bovine Follicle-Stimulating Hormone ß Messenger RNA Downregulates Reporter Expression: Involvement of AU-Rich Elements and Transfactors1

Ravi R. Manjithaya, and Rajan R. Dighe2

Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560012, India

FSHß mRNA has a unique 3' untranslated region (3'UTR) that is highly conserved across the species. Sequence analyses of the mouse, rat, human, bovine, and ovine 3'UTRs revealed the presence of elements implicated in mRNA instability and translational control such as AU-Rich Element (ARE) and lipoxygenase differentiation control elements. Bovine FSHß 3'UTR down-regulated reporter expression in {alpha}T3-1 and NIH3T3 cells, but not in HEK 293 cells, suggesting the involvement of a cell-specific factor or mechanism. The presence of a 3'UTR did not influence reporter mRNA stability, but it did decrease its association with polysomes, indicating that the downregulatory effect may be exerted at the translational level. The segment spanning 601–800 bases (U4) of the bovine FSHß 3'UTR was found to be the most effective downregulating segment, its effect being equal to that of the full-length 3'UTR. RNA electrophoretic mobility shift assay with U4 showed the presence of specific transfactors in the cytosolic preparations of bovine pituitary and the cell lines. U4 contained an ARE that appeared to be functional, because the mutated U4 ARE was ineffective in downregulating the reporter expression and inhibiting [32P]-labeled U4-transfactor complex formation. Downregulation of reporter activity by the full-length 3'UTR and U4 could be overcome by overexpression of HuR, a protein known to stabilize ARE-containing mRNAs in NIH3T3 cells, but not in the {alpha}T3-1 cells, once again indicating that factors other than HuR may also be involved in the regulation of FSHß in the pituitary.

1 Supported by grants from the Indian Council of Medical Research and the University Grant Commission, New Delhi, India.

2 Correspondence. FAX: 91 80 3600999; rdighe{at}mrdg.iisc.ernet.in






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