Journal
INSECT MOLECULAR BIOLOGY
Volume 18, Issue 6, Pages 795-803Publisher
WILEY
DOI: 10.1111/j.1365-2583.2009.00943.x
Keywords
sex determination; gel-shift; UV cross-linking; LC-MS; MS; alternative splicing; Bombyx mori
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Funding
- National Basic Research Program of China [2005CB121000]
- National Hi-Tech Research and Development Program of China [2006AA10A118]
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The Bombyx mori sex determination gene Bmdsx is alternatively spliced in the male and female to produce the male- and female-specific proteins. In an effort to better understand the mechanism of the alternative splicing regulation of Bmdsx, we conducted a gel-shift assay followed by LC-MS/MS analysis to identify the putative proteins bound to the cis-element CE1+6 in the exon 4 of Bmdsx. A protein named as BmHrp28 which is homologous to the Drosophila Hrp48, a member of the hnRNPA/B family, was identified and expressed in Escherichia coli for testing RNA-protein binding in vitro. All of the results showed that BmHrp28 specifically bound to the CE1+6 RNA probe. BmHrp28 has two RNA recognition motifs at the N-terminal and a glycine-rich motif at the C-terminal. It might be one of the factors involved in the male-specific splicing of Bmdsx.
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