Molecular Cloning and Expression Analysis of Bmrbp1 , the Bombyx Mori Homologue of the Drosophila Gene Rbp1

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2009 Sep 5

PMID 19731076

Citations 6

Authors

Zilong Wang

Xingfu Zha

Ningjia He

Zhonghuai Xiang

Qingyou Xa

Affiliations

Soon will be listed here.

Abstract

RBP1 is an important splicing factor involved in alternative splicing of the pre-mRNA of Drosophila sex-determining gene dsx. In this work, the Bombyx mori homologue of the rbp1 gene, Bmrbp1, was cloned. The pre-mRNA of Bmrbp1 gene is alternatively spliced to produce four mature mRNAs, named Bmrbp1-PA, Bmrbp1-PB, Bmrbp1-PC and Bmrbp1-PD, with nucleotide lengths of 799 nt, 1,316 nt, 894 nt and 724 nt, coding for 142 aa, 159 aa, 91 aa and 117 aa, respectively. BmRBP1-PA and BmRBP1-PD contain a N terminal RNA recognization motif (RRM) and a C terminal arginine/serine-rich domain, while BmRBP1-PB and BmRBP1-PC only share a RRM. Amino acid sequence alignments showed that BmRBP1 is conserved with its homologues in other insects and with other SR family proteins. The RT-PCR showed that Bmrbp1-PA was strongly expressed in all examined tissues and development stages, but Bmrbp1-PB was weakly expressed in these tissues and stages. The expression of both Bmrbp1-PA and Bmrbp1-PB showed no obvious sex difference. While the Bmrbp1-PC and Bmrbp1-PD were beyond detection by RT-PCR very likely due to their tissue/stage specificity. These results suggested that Bmrbp1 should be a member of SR family splicing factors, whether it is involved in the sex-specific splicing of Bmdsx pre-mRNA needs further research.

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References

Kanopka A, Muhlemann O, Akusjarvi G . Inhibition by SR proteins of splicing of a regulated adenovirus pre-mRNA. Nature. 1996; 381(6582):535-8. DOI: 10.1038/381535a0. View

Graveley B, Hertel K, Maniatis T . SR proteins are 'locators' of the RNA splicing machinery. Curr Biol. 1999; 9(1):R6-7. DOI: 10.1016/s0960-9822(99)80032-3. View

Norton P . Alternative pre-mRNA splicing: factors involved in splice site selection. J Cell Sci. 1994; 107 ( Pt 1):1-7. DOI: 10.1242/jcs.107.1.1. View

Blencowe B, BOWMAN J, McCracken S, Rosonina E . SR-related proteins and the processing of messenger RNA precursors. Biochem Cell Biol. 1999; 77(4):277-91. View

Suzuki M, Funaguma S, Kanda T, Tamura T, Shimada T . Role of the male BmDSX protein in the sexual differentiation of Bombyx mori. Evol Dev. 2005; 7(1):58-68. DOI: 10.1111/j.1525-142X.2005.05007.x. View

Zahler A, Lane W, Stolk J, Roth M . SR proteins: a conserved family of pre-mRNA splicing factors. Genes Dev. 1992; 6(5):837-47. DOI: 10.1101/gad.6.5.837. View

Kohtz J, Jamison S, Will C, Zuo P, Luhrmann R, Garcia-Blanco M . Protein-protein interactions and 5'-splice-site recognition in mammalian mRNA precursors. Nature. 1994; 368(6467):119-24. DOI: 10.1038/368119a0. View

Andreadis A, Gallego M, Nadal-Ginard B . Generation of protein isoform diversity by alternative splicing: mechanistic and biological implications. Annu Rev Cell Biol. 1987; 3:207-42. DOI: 10.1146/annurev.cb.03.110187.001231. View

Caceres J, Krainer A . Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains. EMBO J. 1993; 12(12):4715-26. PMC: 413916. DOI: 10.1002/j.1460-2075.1993.tb06160.x. View

10.

Breitbart R, Andreadis A, Nadal-Ginard B . Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Annu Rev Biochem. 1987; 56:467-95. DOI: 10.1146/annurev.bi.56.070187.002343. View

11.

Smith C, Valcarcel J . Alternative pre-mRNA splicing: the logic of combinatorial control. Trends Biochem Sci. 2000; 25(8):381-8. DOI: 10.1016/s0968-0004(00)01604-2. View

12.

Bourgeois C, Lejeune F, Stevenin J . Broad specificity of SR (serine/arginine) proteins in the regulation of alternative splicing of pre-messenger RNA. Prog Nucleic Acid Res Mol Biol. 2004; 78:37-88. DOI: 10.1016/S0079-6603(04)78002-2. View

13.

Suzuki M, Ohbayashi F, Mita K, Shimada T . The mechanism of sex-specific splicing at the doublesex gene is different between Drosophila melanogaster and Bombyx mori. Insect Biochem Mol Biol. 2001; 31(12):1201-11. DOI: 10.1016/s0965-1748(01)00067-4. View

14.

Ohbayashi F, Suzuki M, Mita K, Okano K, Shimada T . A homologue of the Drosophila doublesex gene is transcribed into sex-specific mRNA isoforms in the silkworm, Bombyx mori. Comp Biochem Physiol B Biochem Mol Biol. 2001; 128(1):145-58. DOI: 10.1016/s1096-4959(00)00304-3. View

15.

Nagoshi R, McKeown M, Burtis K, Belote J, Baker B . The control of alternative splicing at genes regulating sexual differentiation in D. melanogaster. Cell. 1988; 53(2):229-36. DOI: 10.1016/0092-8674(88)90384-4. View

16.

Wu J, Maniatis T . Specific interactions between proteins implicated in splice site selection and regulated alternative splicing. Cell. 1993; 75(6):1061-70. DOI: 10.1016/0092-8674(93)90316-i. View

17.

Heinrichs V, Baker B . The Drosophila SR protein RBP1 contributes to the regulation of doublesex alternative splicing by recognizing RBP1 RNA target sequences. EMBO J. 1995; 14(16):3987-4000. PMC: 394477. DOI: 10.1002/j.1460-2075.1995.tb00070.x. View

18.

Manley J, Tacke R . SR proteins and splicing control. Genes Dev. 1996; 10(13):1569-79. DOI: 10.1101/gad.10.13.1569. View

19.

Heinrichs V, Baker B . In vivo analysis of the functional domains of the Drosophila splicing regulator RBP1. Proc Natl Acad Sci U S A. 1997; 94(1):115-20. PMC: 19250. DOI: 10.1073/pnas.94.1.115. View

20.

Suzuki M, Funaguma S, Kanda T, Tamura T, Shimada T . Analysis of the biological functions of a doublesex homologue in Bombyx mori. Dev Genes Evol. 2003; 213(7):345-54. DOI: 10.1007/s00427-003-0334-8. View