Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.3.1.28 (chloramphenicol acetyltransferase)
 5,100 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Plant cutin monomers trigger, and glucose suppresses, the expression of the cutinase gene of pathogenic fungi. To identify the cutinase promoter region responsible for induction by the unique plant components, a promoter analysis was done with transformants. Plasmids were constructed that contained (i) the 5' flanking region of the cutinase gene or its deletion mutants from Fusarium solani pisi fused with a chloramphenicol acetyltransferase (CAT) reporter gene and (ii) a constitutive promoter fused with a hygromycin phosphotransferase gene. Hygromycin-resistant transformants of F. solani pisi generated by electroporation were assayed for CAT activity inducible by cutin hydrolysate and for glucose repression of this induction. CAT was induced in a glucose-repressible manner when fused with a 360-base-pair (bp), or longer, segment of the 5' flanking region of the cutinase gene, and deletion of the next 135 bp abolished this induction. Gel retardation assays showed that a protein(s) in nuclear extract from the fungus bound to the 5' flanking region of cutinase gene, and this binding was also abolished when the same 135-bp segment was deleted. These results show that the -225 to -360 segment of the cutinase gene contains a cis-acting regulatory element that binds trans-acting factor(s) in the nuclei. Treatment of the nuclear extract with immobilized phosphatase abolished binding to the promoter, suggesting that binding required a phosphorylated form of the protein. With isolated nuclei, phosphorylation of a protein occurred only in the presence of both cutin monomer and the fungal protein factor. The presence of protein kinase inhibitor H7 during the preincubation of nuclei with the monomer and protein factor inhibited cutinase gene transcription. These results suggest that cutin monomer causes phosphorylation of a transcription factor that binds to the -225 to -360 segment of the cutinase gene and enhances transcription of this gene.
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PMID:Identification of a fungal cutinase promoter that is inducible by a plant signal via a phosphorylated trans-acting factor. 189 70

The cutinase gene from Fusarium solani f. sp. pisi (Nectria haematococca) is induced upon contact with the plant cuticular polymer, cutin, by the unique hydroxy fatty acid monomers released by cutinase carried by virulent strains of the fungus, and this gene is also catabolite-repressed by glucose. Functional elements of the cutinase promoter were studied in vivo by transforming F. solani pisi with fusions of 5'-flanking regions of the cutinase gene and the gene encoding chloramphenicol acetyltransferase (cat). DNA-binding proteins from F. solani pisi were analyzed in vitro by gel shift experiments, methylation interference analysis, and UV-cross-linking experiments. Thus, we identified four promotor elements involved in cutinase gene regulation: a silencer, positive-acting G-rich element, an element that binds a basal transcription factor, and a palindrome necessary for induction by cutin monomer. A silencer between -287 and -249 keeps basal gene expression low but also influences the inducibility of the gene. To restore high levels of induction, a G-rich positive-acting element with sequence similarities to other fungal elements acts as an antagonist to the silencer. Basal transcription is mediated by the first 141 base pairs of the cutinase promoter. The binding site of transcription factor CTF2 was identified between the TATA box and the transcription initiation sites. Gene induction by cutin monomers is regulated by CTF1, most probably a dimeric DNA-binding protein of 49 kDa with a palindromic recognition site at -170.
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PMID:Identification of regulatory elements in the cutinase promoter from Fusarium solani f. sp. pisi (Nectria haematococca). 813 57