Cloning and characterization of hypusine-containing protein eIF5A from the olive flounder Paralichthys olivaceus.
Eukaryotic translation initiation factor 5A (eIF5A) is the only protein in eukaryotic cells that contains the unusual amino acid hypusine (N(epsilon)-(4-amino-2(R)-hydroxybutyl)-lysine). We isolated a 1385-bp eIF5A cDNA containing an open reading frame (ORF) of 468 bp, which encodes a protein of 155 amino acids with a conserved hypusine modification site, from the olive flounder Paralichthys olivaceus. Pairwise alignments revealed that flounder eIF5A had a high sequence identity with those of other known species including mammals. Real-time RT-PCR analysis showed the expression of eIF5A mRNA was constitutively detected in various tissues of healthy flounder. In HINAE cells or flounder kidney infected with the viral hemorrhagic septicemia virus (VHSV), the expression of eIF5A mRNA was slightly increased before cells showed cytopathic effects and then decreased when cells showed cytopathic effects. Treatment of N-guanyl-1,7-diaminoheptane (GC-7), a potent inhibitor of eIF5A hypusination, inhibited the expression of VHSV G protein in a dose-dependent manner suggesting a potential role for eIF5A and its hypusination in viral protein expression.
Kong
Hee Jeong
HJ
Biotechnology Research Institute, National Fisheries Research and Development Institute, 408-1 Sirang-ri, Gijang-up, Gijang-gun, Busan 619-705, Republic of Korea. konghj@nfrdi.go.kr
Hong
Gyeong-Eun
GE
Kim
Woo-Jin
WJ
Kim
Young-Ok
YO
Nam
Bo-Hye
BH
Lee
Chang Hoon
CH
Do
Jeong Wan
JW
Lee
Jeong-Ho
JH
Lee
Sang-Jun
SJ
Kim
Kyung-Kil
KK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005432:Flounder / Q000378:metabolism* / Q000821:virology; D006023:Glycoproteins / Q000096:biosynthesis; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000821:virology; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D008969:Molecular Sequence Data; D029141:Novirhabdovirus / Q000502:physiology; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis; D000097574:Eukaryotic Translation Initiation Factor 5A
D006023:Glycoproteins; D006147:Guanine; D008239:Lysine; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D014764:Viral Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A; C100667:N(1)-guanyl-1,7-diaminoheptane; C100028:hypusine