A DNA methylase from Thermus thermophilus HB8.

A DNA methylase from Thermus thermophilus HB8. Research Abstract Details 

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A DNA methylase from Thermus thermophilus HB8. Abstract Text:

S Sato,K Nakazawa,T Shinomiya,

A DNA methylase was purified in a homogeneous state from a extremely thermophilic bacterium, Thermus thermophilus HB8, by chromatography on, successively, phosphocellulose, CM-cellulose, and heparin-Sepharose. The molecular weight of the enzyme was determined to be about 44,000 by gel filtration on a Sephadex G-100 column and 41,000 by SDS-poly-acrylamide gel electrophoresis, and these findings suggest a single polypeptide enzyme. The enzyme develops maximum activity around pH 7.4 and at 70 degrees C. Enzymatic activity is completely inhibited by 0.2 M NaCl or 2 mM HgCl2. The enzyme transfers methyl groups from S-adenosyl-L-methionine to a double stranded DNA. The sole product of the reaction was identified as N-6-methyl adenine after hydrolysis of the DNA with formic acid. The enzyme kinetics obey the Michaelis-Menten equation and Km values for S-adenosylmethionine and lambda phage DNA were determined to be 0.8 muM and 10 microgram/ml, respectively. The enzyme does not transfer methyl groups to TthHB8I endonuclease digested DNA as well as the host (T. thermophilus HB8) DNA. The number of methyl groups of the fully methylated phiX174 RF DNA was about twice as many as TthHB8I endonuclease sites on the DNA. The distribution of the methyl groups of phiX174 RF DNA among the HaeIII fragments was the same as that of TthHB8I endonuclease sites, suggesting that this DNA methylase is the other component of the modification-restriction system including TthHB8I endonuclease. The enzyme probably recognizes the sequence, 5'-TCGA-3', in a double stranded DNA and probably methylates adenine in the above sequence.

A DNA methylase from Thermus thermophilus HB8. Publishing Authors By Initials

S Sato,K Nakazawa,T Shinomiya,

For similar bacteria: gram-negative bacteria: gram-negative aerobic bacteria: gram-negative aerobic rods and cocci: thermus research abstracts see: bacteria: gram-negative bacteria: gram-negative aerobic bacteria: gram-negative aerobic rods and cocci: thermus research

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A DNA methylase from Thermus thermophilus HB8. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 88

Page Numbers: 737-47

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Sep

YEAR: 1980

A DNA methylase from Thermus thermophilus HB8. Information

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LANGUAGE: eng

NlmUniqueID: 376600

A DNA methylase from Thermus thermophilus HB8. Keywords Mesh Terms:

KEYWORDS: Thermus

MESH TERMS: enzymology

Chemical & Substance for Abstract: A DNA methylase from Thermus thermophilus HB8. Information

Substance Name: DNA (Cytosine-5-)-Methyltransferase

Registry Number: EC 2.1.1.37

Grant and Affiliation Information for A DNA methylase from Thermus thermophilus HB8.

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Country: JAPAN

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MEDLINETA: J Biochem

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