ABSTRACT
Three cDNAs encoding carbonyl reductases (CHC1-3) have been isolated and expressed in bacterial cells. The recombinant enzymes were further characterized with respect to substrate specificity, inhibitor sensitivity and response to pyruvate. In addition, the expression of the mRNAs of CHCRs encoding in brain, liver and kidney was analyzed by RT-PCR. Searches of EST files revealed that orthologues of both CHCR3 and human CBR3 are expressed in variety of human cells and tissues.
Subject(s)
Alcohol Oxidoreductases/metabolism , Alcohol Oxidoreductases/antagonists & inhibitors , Alcohol Oxidoreductases/chemistry , Alcohol Oxidoreductases/genetics , Amino Acid Sequence , Animals , Base Sequence , Cricetinae , Cricetulus , Enzyme Inhibitors/pharmacology , Expressed Sequence Tags , Humans , Models, Molecular , Molecular Sequence Data , RNA, Messenger/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Substrate SpecificityABSTRACT
Using cDNA-based array analysis combined with double-stranded RNA interference (dsRNAi), we have identified yk298h6 as a target gene of Caenorhabditis elegans TGF-beta signaling. Worms overexpressing dbl-1, a TGF-beta ligand, are 16% longer than wild type. Array analysis shows yk298h6 to be one of several genes suppressed in such worms. Disruption of yk298h6 function by dsRNAi also resulted in long worms, suggesting that it is a negative regulator of body length. yk298h6 was then mapped to, and shown to be identical to, lon-1, a known gene that affects body length. lon-1 encodes a 312 amino acid protein with a motif sequence that is conserved from plants to humans. Expression studies confirm that LON-1 is repressed by DBL-1, suggesting that LON-1 is a novel downstream component of the C.elegans TGF-beta growth regulation pathway. Consistent with this, LON-1 is expressed mainly in the larval and adult hypodermis and has dose-dependent effects on body length associated with changes in hypodermal ploidy, but not hypodermal cell proliferation.
