Targeted disruption of the genes, mlcR and ariB, which encode GAL4-type proteins in Penicillium citrinum.
The role of two genes, mlcR and ariB, was investigated by gene disruption experiments. The mlcR gene in the ML-236B biosynthetic gene cluster of Penicillium citrinum encodes a putative 50.2-kDa protein with a Zn (II) 2Cys6 DNA-binding domain, and has similarity to most of the GAL4-type regulatory proteins. The mlcR disruptant did not produce ML-236B or its intermediates, suggesting that mlcR is involved in ML-236B biosynthesis. Transcriptional analysis of the mlcR disruptant by Northern hybridization and RT-PCR indicated that MlcR activates the transcription of mlcA, B, C,D, F, G and H in a pathway-specific manner. On the other hand, MlcR did not affect the transcription of mlcE and the genes outside the ML-236B cluster. The ariB gene, next to mlcR, encodes another GAL4-type protein. Transcriptional analysis of the ariB disruptant indicated that it is a transcriptional activator of the genes outside the ML-236B cluster, and is not related to ML-236B biosynthesis.
Baba
S
S
Process Development Laboratories, Sankyo Co., Ltd., Fukushima, Japan. satoshib@sankyo.co.jp
Abe
Y
Y
Ono
C
C
Hosobuchi
M
M
D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018390:Gene Targeting; D005800:Genes, Fungal; D005822:Genetic Vectors; D005810:Multigene Family; D010407:Penicillium / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005656:Fungal Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors