Expression and characterization of the Streptomyces coelicolor serine/threonine protein kinase PkaD.
We identified and characterized the gene encoding a new eukaryotic-type protein kinase from Streptomyces coelicolor A3(2) M145. PkaD, consisting of 598 amino acid residues, contained the catalytic domain of eukaryotic protein kinases in the N-terminal region. A hydrophobicity plot indicated the presence of a putative transmembrane spanning sequence downstream of the catalytic domain, suggesting that PkaD is a transmembrane protein kinase. The recombinant PkaD was found to be phosphorylated at the threonine and tyrosine residues. In S. coelicolor A3(2), pkaD was transcribed as a monocistronic mRNA, and it was expressed constitutively throughout the life cycle. Disruption of chromosomal pkaD resulted in a significant loss of actinorhodin production. This result implies the involvement of pkaD in the regulation of secondary metabolism.
Urabe
Hiroaki
H
Department of Biochemistry, Meiji Pharmaceutical University, Kiyose, Tokyo 204-8588, Japan. hurabe@my-pharm.ac.jp
Aoyagi
Narumi
N
Ogawara
Hiroshi
H
Motojima
Kiyoto
K
D000880:Anthraquinones / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D020134:Catalytic Domain; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins; D008660:Metabolism; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D048372:Streptomyces coelicolor; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D000095622:Benzoisochromanequinones
D000880:Anthraquinones; D001426:Bacterial Proteins; D008565:Membrane Proteins; D017346:Protein Serine-Threonine Kinases; D014157:Transcription Factors; C092410:PkaA protein, Streptomyces coelicolor; C013390:actinorhodin; D000095622:Benzoisochromanequinones