Identification of low-molecular-weight vitellogenin 1 (Vg1)-like proteins as nucleotide excision repair (NER) factors in developing zebrafish (Danio rerio) using a transcription-based DNA repair assay

Abstract

Nucleotide excision repair (NER) removes helix-distorting DNA lesions such as UV-induced pyrimidine dimers and cisplatin-induced strand crosslinking. Our earlier studies have identified low-molecular-weight proteins homologous to the 150-kDa vitellogenin 1 (Vg1) as UV-damaged DNA-binding factors expressed in developing zebrafish (Danio rerio). This present study explored if Vg1-like proteins also participated in NER in zebrafish. Immunoblot analysis of affinity-captured 12 h post-fertilization (hpf) zebrafish extract proteins showed a transient binding of a 30-kDa Vg1-like polypeptide to UV-damaged DNA. A transcription-based in vitro repair assay revealed a significant up-regulation of UVC or cisplatin-suppressed transcriptional activity of a marker cDNA driven by a SP6 RNA polymerase-regulated promotor after incubating the damaged plasmid with the extracts of 12 hpf embryos or 96 hpf larvae. The up-regulation of UV or cisplatin-suppressed transcription was abolished in the presence of a monoclonal anti-zebrafish Vg1 antibody. The differential sensitivity of UV-induced repair in 12 and 96 hpf zebrafish extracts to exogenous ATP suggested a development-dependent expression of Vg1-like NER factors. A T endonuclease V digestion assay showed no inhibition of the anti-Vg1 antibody on the excision of UV-induced cyclobutane pyrimidine dimers. Our results identified the participation of Vg1-like factors in NER in developing zebrafish, and these factors may function at post-incison steps of NER.

The ZcVgR gene is involved in the regulation of reproduction and lifespan in female Zeugodacus cucurbitae (Coquillett) mediated by short-term high temperature

“…Compared to the transcriptional level, the number of differential proteins related to reproduction and lifespan was low, with ZcVg3, Vg1-like, HSP23like and GPCR Mth2-like genes signi cantly up-regulated and TnC genes signi cantly downregulated at both transcriptional and protein levels. The HSP23 gene is mainly associated with diapause of insects and is upregulated during diapause of pupa overwintering in Sarcophaga crassipalpis (Rinehart et al, 2007), the GPCR Mth2 gene plays a role in drug resistance in Musca domestica (Ma et al, 2020), and the Vg1-like gene is involved in nucleic acid excision during the developmental repair process in Danio rerio (Shen et al, 2017). The Vg1-like, HSP23-like, and GPCR Mth2-like genes of female Z. cucurbitae may have similar functions, but further studies are needed.…”

DNA repair genes play a variety of roles in the development of fish embryos

“…The Xpa protein expression was not detected until 84 hpf. Later, Shen et al (2017) confirmed that the vitellogenin-1-like protein in zebrafish embryos plays a role of UV-damaged-DNA binding factor and revealed its function in postincision step of NER (Shen et al, 2017).…”