cDNA cloning and characterization of human Delta5-desaturase involved in the biosynthesis of arachidonic acid.
Two human expressed sequence tag (EST) cDNA sequences with identity with Delta(5)- and Delta(6)-desaturases from a filamentous fungus, Mortierella alpina, were identified from the LifeSeq(R) database of Incyte Pharmaceuticals, Inc. (Palo Alto, CA, U.S.A.). An oligonucleotide complementary to the 3' EST cDNA sequences was used to screen human liver cDNA using rapid amplification of cDNA ends (RACE)-PCR. The amplified DNA fragment had 98% identity with a putative open reading frame (ORF) predicted from a human genomic sequence, and encoded 444 amino acids. Expression of this ORF in mouse fibroblast cells demonstrated that the encoded protein was a Delta(5)-desaturase, as determined by the conversion of dihomo-gamma-linolenic acid (C(20:3,n-6)) into arachidonic acid (C(20:4,n-6)). The human Delta(5)-desaturase contained a predicted N-terminal cytochrome b(5)-like domain, as well as three histidine-rich domains. A tissue expression profile revealed that this gene is highly expressed in fetal liver, fetal brain, adult brain and adrenal gland. A search of the existing databases led to localization of this ORF within a 14 kb interval flanked by the flap endonuclease-1 (FEN1) and vitelliform macular dystrophy (Best's disease; VMD2) loci of chromosome 11q12.
Leonard
A E
AE
Department of Strategic Discovery Research, Ross Products Division, Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, USA.
Kelder
B
B
Bobik
E G
EG
Chuang
L T
LT
Parker-Barnes
J M
JM
Thurmond
J M
JM
Kroeger
P E
PE
Kopchick
J J
JJ
Huang
Y S
YS
Mukerji
P
P
D015126:8,11,14-Eicosatrienoic Acid / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016718:Arachidonic Acid / Q000096:biosynthesis* / Q000378:metabolism; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D016208:Databases, Factual; D000090943:Delta-5 Fatty Acid Desaturase; D020224:Expressed Sequence Tags; D044943:Fatty Acid Desaturases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005227:Fatty Acids / Q000032:analysis; D020869:Gene Expression Profiling; D006801:Humans; D007739:L Cells; D050602:Linoleoyl-CoA Desaturase; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D020161:Physical Chromosome Mapping; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014162:Transfection; D017965:gamma-Linolenic Acid / Q000378:metabolism
D018076:DNA, Complementary; D000090943:Delta-5 Fatty Acid Desaturase; D005227:Fatty Acids; D012333:RNA, Messenger; D016718:Arachidonic Acid; D017965:gamma-Linolenic Acid; D044943:Fatty Acid Desaturases; D050602:Linoleoyl-CoA Desaturase; D015126:8,11,14-Eicosatrienoic Acid