Molecular cloning and stress-induced expression of paralichthys olivaceus heme-regulated initiation factor 2alpha kinase.
The heme-regulated initiation factor 2alpha kinase (HRI) is acknowledged to play an important role in translational shutoff in reticulocytes in response to various cellular stresses. In this study, we report its homologous cDNA cloning and characterization from cultured flounder embryonic cells (FEC) after treatment with UV-inactivated grass carp haemorrhagic virus (GCHV). The full-length cDNA of Paralichthys olivaceus HRI homologue (PoHRI) has 2391 bp and encodes a protein of 651 amino acids. The putative PoHRI protein exhibits high identity with all members of eIF2alpha kinase family. It contains 12 catalytic subdomains located within the C-terminus of all Ser/Thr protein kinases, a unique kinase insertion of 136 amino acids between subdomains IV and V, and a relatively conserved N-terminal domain (NTD). Upon heat shock, virus infection or Poly I:C treatment, PoHRI mRNA and protein are significantly upregulated in FEC cells but show different expression patterns in response to different stresses. In healthy flounders, PoHRI displays a wide tissue distribution at both the mRNA and protein levels. These results indicate that PoHRI is a ubiquitous eIF2alpha kinase and might play an important role in translational control over nonheme producing FEC cells under different stresses.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate School of Chinese Academy of Sciences, Wuhan 430072, China.
Rong
R
Zhu
Yi-Bing
YB
Zhang
Yu-Dong
YD
Chen
Cai-Wen
CW
Dong
Fu-Tie
FT
Zhang
Qi-Ya
QY
Zhang
Jian-Fang
JF
Gui
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005432:Flounder / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D018869:Heat-Shock Response / Q000276:immunology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011070:Poly I-C / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D013312:Stress, Physiological / Q000201:enzymology / Q000276:immunology / Q000662:veterinary*; D019892:eIF-2 Kinase / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D012333:RNA, Messenger; D019892:eIF-2 Kinase; D011070:Poly I-C