Cloning and analysis of the gene for cystathionine gamma-synthase from Arabidopsis thaliana.
A cDNA clone, CGS1, encoding cystathionine gamma-synthase (CGS) from Arabidopsis thaliana was selected by complementation of CGS mutant strain of Escherichia coli (metB). Cells expressing CGS1 can grow on medium lacking Met and contain CGS enzyme activity. Genomic DNA blot analysis of A. thaliana revealed that there is a single gene homologous with CGS1. A genomic fragment carrying CGS1 was cloned and sequenced. Through combined analysis of the cDNA and genomic clone it was determined that the CGS1 coding sequence is 1692 bp, encodes a 563 amino acid, 60 kDa protein, and is interrupted by ten introns. A transcriptional initiation site was detected 260 bp 5' of the initiator codon. The predicted amino acid sequence of CGS1 contains a consensus pyridoxal phosphate-binding site and is similar to MetB of E. coli, with which it is 35 percent identical. The CGS1 product has a sequence at the amino terminus that resembles a transit peptide for localization to plastids. At least 160 amino acids from the amino terminus of the CGS1 enzyme are not essential for enzymatic activity.
Kim
J
J
Center for Agricultural Molecular Biology and Department of Food Science, Rutgers University, New Brunswick, NJ 08903-0231, USA.
Leustek
T
T
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D019757:Carbon-Oxygen Lyases; D003001:Cloning, Molecular; D018387:Codon, Initiator; D018076:DNA, Complementary / Q000235:genetics; D017343:Genes, Plant; D015698:Genomic Library; D008190:Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011732:Pyridoxal Phosphate / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D018387:Codon, Initiator; D018076:DNA, Complementary; D011732:Pyridoxal Phosphate; C022696:O-succinylhomoserine (thiol)-lyase; D008190:Lyases; D019757:Carbon-Oxygen Lyases