☆ 4.5 Article

Characterization of nuclear localization signal in Ostrinia furnacalis Masculinizer protein

ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (2021)

Journal

ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY

Volume 106, Issue 3, Pages -

Publisher

WILEY

DOI: 10.1002/arch.21768

Keywords

bipartite NLS; Bombyx mori; masculinization; Masculinizer protein; Ostrinia furnacalis

Funding

Japan Society for the Promotion of Science [17H06431]
Grants-in-Aid for Scientific Research [17H06431] Funding Source: KAKEN

Ask authors/readers for more resources

Protocol

Community support

Reagent

Community support

Automated Summary New
Abstract

The Bombyx mori Masculinizer protein (BmMasc) and Ostrinia furnacalis Masc (OfMasc) both have nuclear localization signals, but their functional properties and critical residues differ; Masc proteins function in the cytoplasm to drive the masculinizing cascade.

Bombyx mori Masculinizer protein (BmMasc) is essential for both masculinization and dosage compensation in B. mori. We previously identified a bipartite nuclear localization signal (NLS) of BmMasc and two essential residues (lysine at 274 [K274] and arginine at 275 [R275]) implicated in its function. Sequence comparison showed the presence of putative NLSs in lepidopteran Masc proteins, but their functional properties and critical residues are unknown. Here we characterized a putative NLS of Ostrinia furnacalis Masc (OfMasc) using B. mori ovary-derived BmN-4 cell line. Deletion and alanine scanning mutagenesis revealed that a putative NLS is required for nuclear localization of OfMasc. However, mutations at both K227 and R228, which correspond to K274 and R275 of BmMasc, respectively, do not greatly abolish the NLS activity. Additional mutagenesis analysis revealed that triple mutations at K227, R228, and K240 almost completely inhibited OfMasc nuclear localization. These results suggest that lepidopteran Masc proteins possess a common functional NLS, but the critical residues for its activity are different. Moreover, we examined the masculinizing activity of OfMasc derivatives and found that nuclear localization is not required for the masculinizing activity of OfMasc. The results from our studies indicate that lepidopteran Masc proteins function in the cytoplasm to drive masculinizing cascade.

Authors

I am an author on this paper

Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.5

Not enough ratings

Secondary Ratings

Novelty

Significance

Scientific rigor

Rate this paper

Masculinizer is not post-transcriptionally regulated by female-specific piRNAs during sex determination in the Asian corn borer, Ostrinia furnacalis

Takahiro Fukui, Keisuke Shoji, Takashi Kiuchi, Yutaka Suzuki, Susumu Katsuma

Summary: Lepidopteran insects show a different mechanism of sex determination compared to most insect species. The silkworm Bombyx mori, a model species in this group, has a sex-determining gene called Feminizer (Fem) on the female-specific W chromosome. Fem is a precursor of PIWI-interacting small RNA (piRNA) that cleaves the mRNA of the male-determining gene Masc, leading to female development. However, this piRNA-dependent sex determination strategy is not observed in the Asian corn borer, Ostrinia furnacalis. Despite previous findings of masculinizing function of the Masc gene (OfMasc) in O. furnacalis embryos, there was no difference in the expression level of OfMasc between males and females during sex determination. Furthermore, deep sequencing analysis did not identify any female-specific small RNAs mapped onto OfMasc mRNA. Knockdown of two PIWI genes also did not affect the expression level of OfMasc in either sex. These findings suggest the possibility of divergent evolution of sex determinants in Lepidoptera.

INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY (2023)