Molecular cloning and expression of a rat cDNA encoding 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase.
The cDNA of a 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (AICARFT/IMPCHase) was isolated from rat liver RNA by reverse transcription and the polymerase chain reaction (PCR). The rat AICARFT/IMPCHase cDNA included 1928 bp containing a coding region of 1779 bp for a 592-amino acid polypeptide (Mr = 64 200). Rat and human AICARFT/IMPCHase cDNAs show 84 and 91% homology at the nucleotide and amino acid sequence level, respectively. The protein produced by the rat cDNA using pET-expression system catalysed the penultimate and final steps of de novo purine biosynthesis. Northern analysis identified a 2.8-kb AICARFT/IMPCHase mRNA and the level of the AICARFT/IMPCHase transcripts increased markedly at 24 h after partial (70%) hepatectomy.
Akira
T
T
Department of Food Science and Nutrition, Nara Women's University, Japan.
Komatsu
M
M
Nango
R
R
Tomooka
A
A
Konaka
K
K
Yamauchi
M
M
Kitamura
Y
Y
Nomura
S
S
Tsukamoto
I
I
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D006498:Hepatectomy; D019877:Hydroxymethyl and Formyl Transferases / Q000235:genetics*; D008099:Liver / Q000502:physiology; D008115:Liver Regeneration / Q000235:genetics; D008969:Molecular Sequence Data; D009709:Nucleotide Deaminases / Q000235:genetics*; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019877:Hydroxymethyl and Formyl Transferases; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D009709:Nucleotide Deaminases; C021436:IMP cyclohydrolase