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Characterization of CgHIF alpha-Like, a Novel bHLH-PAS Transcription Factor Family Member, and Its Role under Hypoxia Stress in the Pacific Oyster Crassostrea gigas
Wang, Ting1,2,3,5; Meng, Jie1,4,5; Li, Li1,4,5; Zhang, Guofan1,3,5
2016-11-04
Source PublicationPLOS ONE
Volume11Issue:11
SubtypeArticle
AbstractHypoxia-inducible factor (HIF), a critical member of the basic-helix-loop-helix (bHLH)-containing Per-Arnt-Sim (PAS) protein family, is a master transcription factor involved in maintaining oxygen homeostasis. In the present study, we isolated and characterized a novel bHLH-PAS family member, CgHIF alpha-like gene, from the Pacific oyster Crassostrea gigas, and determined its importance during hypoxia stress. The 3020-bp CgHIF alpha-like cDNA encoded a protein of 888 amino acids. The predicted CgHIF alpha-like amino acid sequence was conserved in the N-terminal bHLH, PAS, and PAC domains (but not in the C-terminal domain) and was most closely related to the HIF family in the bHLH-PAS protein phylogenic tree. Similar to the mammalian HIF-1a, CgHIF alpha-like could be expressed as four mRNA iso-forms containing alternative 50-untranslated regions and different translation initiation codons. At the mRNA level, these isoforms were expressed in a tissue-specific manner and showed increased transcription to varying degrees under hypoxic conditions. Additionally, the western blot analysis demonstrated that CgHIF alpha-like was induced by hypoxia. Electrophoretic mobility shift assay indicated that CgHIF alpha-like could bind to the hypoxia responsive element (HRE), whereas dual-luciferase reporter analysis demonstrated that CgHIF alpha-like could transactivate the reporter gene containing the HREs. In addition to CgHIF alpha-like, we identified CgARNT from the C. gigas, analyzed its expression pattern, and confirmed its interaction with CgHIF alpha-like using a yeast two-hybrid assay. In conclusion, this is the first report on the cloning and characterization of a novel hypoxia transcription factor in mollusks, which could accumulate under hypoxia and regulate hypoxia related gene expression by binding to HRE and dimerizing with CgARNT. As only one member of HIF has been identified in invertebrates to date, our results provide new insights into the unique mechanisms of hypoxia tolerance in mollusks.
DOI10.1371/journal.pone.0166057
Indexed BySCI
Language英语
WOS IDWOS:000386911100028
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.qdio.ac.cn/handle/337002/136201
Collection海洋生物技术研发中心
实验海洋生物学重点实验室
Affiliation1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
4.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries & Aquaculture, Qingdao, Shandong, Peoples R China
5.Chinese Acad Sci, Natl & Local Joint Engn Lab Ecol Mariculture, Inst Oceanol, Qingdao, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Wang, Ting,Meng, Jie,Li, Li,et al. Characterization of CgHIF alpha-Like, a Novel bHLH-PAS Transcription Factor Family Member, and Its Role under Hypoxia Stress in the Pacific Oyster Crassostrea gigas[J]. PLOS ONE,2016,11(11).
APA Wang, Ting,Meng, Jie,Li, Li,&Zhang, Guofan.(2016).Characterization of CgHIF alpha-Like, a Novel bHLH-PAS Transcription Factor Family Member, and Its Role under Hypoxia Stress in the Pacific Oyster Crassostrea gigas.PLOS ONE,11(11).
MLA Wang, Ting,et al."Characterization of CgHIF alpha-Like, a Novel bHLH-PAS Transcription Factor Family Member, and Its Role under Hypoxia Stress in the Pacific Oyster Crassostrea gigas".PLOS ONE 11.11(2016).
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