PimM, a PAS domain positive regulator of pimaricin biosynthesis in Streptomyces natalensis.
Sequencing of the DNA region on the left fringe of the pimaricin gene cluster revealed the presence of a 579 bp gene, pimM, whose deduced product (192 aa) was found to have amino acid sequence homology with bacterial regulatory proteins. Database comparisons revealed that PimM combines an N-terminal PAS domain with a C-terminal helix-turn-helix (HTH) motif of the LuxR type. Gene replacement of pimM from the Streptomyces natalensis chromosome with a mutant version lacking the HTH DNA-binding domain resulted in complete loss of pimaricin production, suggesting that PimM is a positive regulator of pimaricin biosynthesis. Complementation of the DeltapimM mutant with a single copy of pimM integrated into the chromosome restored pimaricin production. The insertion of a single copy of pimM, with its own promoter, into the S. natalensis wild-type strain boosted pimaricin production. Gene expression analyses in S. natalensis wild-type and DeltapimM by reverse transcriptase PCR (RT-PCR) of the pimaricin gene cluster revealed the targets for the PimM regulatory protein. According to these analyses, the genes responsible for initiation and first elongation cycles of polyketide chain extension are among the major targets for regulation. Other pim genes are differentially affected. Interestingly, our results indicate that PimM plays its regulatory role independently of PimR, the first pathway-specific regulator of pimaricin biosynthesis.
Antón
Nuria
N
Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.|Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
Santos-Aberturas
Javier
J
Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.|Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
Mendes
Marta V
MV
Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
Guerra
Susana M
SM
Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.|Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
Martín
Juan F
JF
Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.|Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
Aparicio
Jesús F
JF
Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.|Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real no 1, 24006 León, Spain.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D019077:Helix-Turn-Helix Motifs; D008969:Molecular Sequence Data; D010866:Natamycin / Q000096:biosynthesis*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; C064778:LuxR autoinducer binding proteins; D010866:Natamycin