title: Identification of the gene for disaggregatase from Methanosarcina mazei.
abstract: The gene sequences encoding disaggregatase (Dag), the enzyme responsible for dispersion of cell aggregates of Methanosarcina mazei to single cells, were determined for three strains of M. mazei (S-6(T), LYC and TMA). The dag genes of the three strains were 3234 bp in length and had almost the same sequences with 97% amino acid sequence identities. Dag was predicted to comprise 1077 amino acid residues and to have a molecular mass of 120 kDa containing three repeats of the DNRLRE domain in the C terminus, which is specific to the genus Methanosarcina and may be responsible for structural organization and cell wall function. Recombinant Dag was overexpressed in Escherichia coli and preparations of the expressed protein exhibited enzymatic activity. The RT-PCR analysis showed that dag was transcribed to mRNA in M. mazei LYC and indicated that the gene was expressed in vivo. This is the first time the gene involved in the morphological change of Methanosarcina spp. from aggregate to single cells has been identified.
authors.affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Osumi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kakehashi
authors.forename: Shiho
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Kazunari
authors.initials: K
authors.lastname: Nagaoka
authors.forename: Junichi
authors.initials: J
authors.lastname: Sakai
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Miyashita
authors.forename: Makoto
authors.initials: M
authors.lastname: Kimura
authors.forename: Susumu
authors.initials: S
authors.lastname: Asakawa
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D019848:Gene Expression Regulation, Archaeal; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017020:Methanosarcina / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
chemical_list: D019843:Archaeal Proteins; C067993:DAG protein, Methanosarcina mazei; D006026:Glycoside Hydrolases