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P-element Somatic Inhibitor Protein Binding a Target Sequence in dsx Pre-mRNA Conserved in Bombyx mori and Spodoptera litura

Authors
  • Wang, Yao1
  • Zhao, Qin1
  • Wan, Qiu-Xing1
  • Wang, Kai-Xuan1
  • Zha, Xing-Fu1, 2, 3
  • 1 (K.-X.W.)
  • 2 Chongqing Key Laboratory of Sericultural Science, Southwest University, Chongqing 400715, China
  • 3 Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Chongqing 400715, China
Type
Published Article
Journal
International Journal of Molecular Sciences
Publisher
MDPI AG
Publication Date
May 13, 2019
Volume
20
Issue
9
Identifiers
DOI: 10.3390/ijms20092361
PMID: 31086020
PMCID: PMC6539025
Source
PubMed Central
Keywords
License
Green

Abstract

Bombyx mori doublesex ( Bmdsx ) functions as a double-switch gene in the final step of the sex-determination cascade in the silkworm Bombyx mori . The P-element somatic inhibitor (PSI) protein in B. mori interacts with Bmdsx pre-mRNA in CE1 as an exonic splicing silencer to promote male-specific splicing of Bmdsx . However, the character of the interaction between BmPSI and Bmdsx pre-mRNA remains unclear. Electrophoretic mobility shift assay (EMSA) results showed that the four KH_1 motifs in BmPSI are all essential for the binding, especially the former two KH_1 motifs. Three active sites (I116, L127, and IGGI) in the KH_1 motif were found to be necessary for the binding through EMSA, circular dichroism (CD) spectroscopy, and isothermal titration calorimetry (ITC). The PSI homologous protein in S. litura (SlPSI) was purified and the binding of SlPSI and CE1 was verified. Compared with BmPSI, the mutant SlPSI proteins of I116 and IGGI lost their ability to bind to CE1. In conclusion, the binding of PSI and dsx pre-mRNA are generally conserved in both B. mori and S. litura . These findings provide clues for sex determination in Lepidoptera.

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