title: Characterization of the autophosphorylating kinase, PkaF, in Streptomyces coelicolor A3(2) M130.
abstract: Streptomyces coelicolor, the model species for morphologically complex actinomycete bacteria, has unique characteristics such as morphological and physiological differentiation, which are controlled by various factors and several protein kinases. From the whole genomic sequence of S. coelicolor A3(2), 44 putative serine/threonine (Ser/Thr) protein kinases were identified, and the pkaF gene was chosen as the best-conserved protein for typical Ser/Thr protein kinases. pkaF encodes a 667-amino acid protein with a predicted N-terminal Ser/Thr kinase domain and four repeated C-terminal penicillin-binding domains and Ser/Thr kinase-associated (PASTA) domains. Based on PCR, a pkaF gene was cloned and heterologously expressed. PkaF expressed in Escherichia coli had the bigger molecular size than the expected value (75 kDa) and was further purified by Ni2+-NTA agarose affinity column chromatography to homogeneity. The purified PkaF was autophosphorylated through the transfer of the γ-phosphate group of ATP. The extent of phosphorylation was proportional to the amount of PkaF, and the phospho-PkaF was dephosphorylated by the addition of the cell lysate of S. coelicolor A3(2). Although no change was observed in the pkaF disruptant, overexpression of pkaF induced severe repression of morphogenesis and actinorhodin production, but not undecylprodigiosin production, implying that PkaF specifically regulates morphogenesis and actinorhodin production in S. coelicolor.
authors.affiliation: Department of Biological Science and Institute of Bioscience and Biotechnology, Myongji University, Yongin, 449-728, Korea.
authors.forename: Eun A
authors.initials: EA
authors.lastname: Oh
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Chi
authors.forename: Mi-Soon
authors.initials: MS
authors.lastname: Kim
authors.forename: Sang Sun
authors.initials: SS
authors.lastname: Kang
authors.forename: Jaesun
authors.initials: J
authors.lastname: Chun
authors.forename: Soon-Kwang
authors.initials: SK
authors.lastname: Hong
mesh_terms: D000595:Amino Acid Sequence; D000880:Anthraquinones / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000201:enzymology* / Q000235:genetics; D000095622:Benzoisochromanequinones
chemical_list: D000880:Anthraquinones; D001426:Bacterial Proteins; D017346:Protein Serine-Threonine Kinases; C013390:actinorhodin; D000095622:Benzoisochromanequinones