Alternative splicing transcription of Megalobrama amblycephala HIF prolyl hydroxylase PHD3 and up-regulation of PHD3 by HIF-1α

Nan Chen; Cui-Hong Huang; Bo-Xiang Chen; Hong Liu; Wei-Min Wang; Yasmeen Gul; Huan-Ling Wang

doi:10.1016/j.bbrc.2015.12.046

Alternative splicing transcription of Megalobrama amblycephala HIF prolyl hydroxylase PHD3 and up-regulation of PHD3 by HIF-1α

Biochem Biophys Res Commun. 2016 Jan 15;469(3):737-42. doi: 10.1016/j.bbrc.2015.12.046. Epub 2015 Dec 15.

Authors

Nan Chen¹, Cui-Hong Huang¹, Bo-Xiang Chen², Hong Liu³, Wei-Min Wang¹, Yasmeen Gul⁴, Huan-Ling Wang⁵

Affiliations

¹ Key Lab of Freshwater Animal Breeding, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070, Wuhan, PR China.
² Key Lab of Freshwater Animal Breeding, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070, Wuhan, PR China; Hubei BaiRong Improved Aquatic Seed Co., Ltd, 438800 Huanggang, PR China.
³ Key Lab of Freshwater Animal Breeding, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070, Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070, Wuhan, PR China.
⁴ Department of Zoology, Government College Women University Faisalabad, Pakistan.
⁵ Key Lab of Freshwater Animal Breeding, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070, Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070, Wuhan, PR China. Electronic address: hbauwhl@hotmail.com.

PMID: 26697748
DOI: 10.1016/j.bbrc.2015.12.046

Abstract

PHD3 is a hydroxylase that hydroxylates prolyl residues on hypoxia-inducible factors (HIFs) in mammals. In this study, the full-length cDNA and promoter sequences of Megalobrama amblycephala PHD3 gene were isolated by a modified RACE method. PHD3 cDNA was 1622 bp in length, including an ORF of 717 bp encoding 238 amino acid residues. The semi-quantitative PCR results suggested that PHD3 was highly expressed in liver in the normal condition, while after hypoxia treatment this gene was significantly increased in all analyzed tissues. PHD3 was detected only in the initial stages of M. amblycephala embryo development. In addition, the presence of another alternatively processed PHD3 transcript, designated PHD3Δ1 was observed in the process of analyzing the expression of PHD3. Both PHD3 and PHD3Δ1 were up-regulated under hypoxia, and had five the hypoxia response elements (HREs) by in silico scanning on the promoter. Further luciferase assay indicated that all HREs significantly responded to hypoxia. Taken together, these results suggest that PHD3 plays important roles in hypoxia response and early embryo development of M. amblycephala.

Keywords: Alternative splicing transcription; Hypoxia; Megalobrama amblycephala; PHD3.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alternative Splicing / genetics*
Animals
Embryonic Development / genetics*
Fishes / genetics*
Gene Expression Regulation, Developmental / genetics
Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
Hypoxia-Inducible Factor-Proline Dioxygenases / genetics*
Transcriptional Activation / genetics*
Up-Regulation / genetics

Substances

HIF1A protein, human
Hypoxia-Inducible Factor 1, alpha Subunit
Hypoxia-Inducible Factor-Proline Dioxygenases