Identification of the GGPS1 genes encoding geranylgeranyl diphosphate synthases from mouse and human.
E,E,E-Geranylgeranyl diphosphate (GGPP) is an important precursor of carotenoids and geranylgeranylated proteins such as small G proteins. In this study, we have identified mouse and human GGPP synthase genes. Sequence analysis showed that mouse and human GGPP synthases share a high level of amino acid identity (94%) with each other, and share a high level of similarity (45-50%) with GGPP synthases of lower eukaryotes, but only weak similarity (22-31%) to plant and prokaryotic GGPP synthases. Both of the newly identified GGPP synthase genes from mouse and human were expressed in Escherichia coli, and their gene products displayed GGPP synthase activity when isopentenyl diphosphate and farnesyl diphosphate were used as substrates. The GGPP synthase activity of these genes was also confirmed by demonstrating carotenoid synthesis after co-transformation of E. coli with a plasmid expressing the crt genes derived from Erwinia uredovora, and a plasmid expressing either the mouse or human GGPS1 gene. Southern blot analysis suggests that the human GGPS1 gene is a single copy gene.
Kainou
T
T
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue 690-8504, Japan.
Kawamura
K
K
Tanaka
K
K
Matsuda
H
H
Kawamukai
M
M
D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D004122:Dimethylallyltranstransferase; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D051231:Farnesyltranstransferase; D051228:Geranyltranstransferase; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D010802:Phylogeny; D010957:Plasmids; D016415:Sequence Alignment
C495428:Ggps1 protein, mouse; D009097:Multienzyme Complexes; D019883:Alkyl and Aryl Transferases; D004122:Dimethylallyltranstransferase; D051228:Geranyltranstransferase; D051231:Farnesyltranstransferase; C495427:GGPS1 protein, human