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PMID: 2690017 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Purification, cloning and sequence analysis of RsrI DNA methyltransferase: lack of homology between two enzymes, RsrI and EcoRI, that methylate the same nucleotide in identical recognition sequences.

Nucleic acids research ·Vol. 17 ·No. 24 ·1989-12-25 ·Pages 10403-25

Kaszubska W, Aiken C, O'Connor CD, Gumport RI

Abstract

RsrI DNA methyltransferase (M-RsrI) from Rhodobacter sphaeroides has been purified to homogeneity, and its gene cloned and sequenced. This enzyme catalyzes methylation of the same central adenine residue in the duplex recognition sequence d(GAATTC) as does M-EcoRI. The reduced and denatured molecular weight of the RsrI methyltransferase (MTase) is 33,600 Da. A fragment of R. sphaeroides chromosomal DNA exhibited M.RsrI activity in E. coli and was used to sequence the rsrIM gene. The deduced amino acid sequence of M.RsrI shows partial homology to those of the type II adenine MTases HinfI and DpnA and N4-cytosine MTases BamHI and PvuII, and to the type III adenine MTases EcoP1 and EcoP15. In contrast to their corresponding isoschizomeric endonucleases, the deduced amino acid sequences of the RsrI and EcoRI MTases show very little homology. Either the EcoRI and RsrI restriction-modification systems assembled independently from closely related endonuclease and more distantly related MTase genes, or the MTase genes diverged more than their partner endonuclease genes. The rsrIM gene sequence has also been determined by Stephenson and Greene (Nucl. Acids Res. (1989) 17, this issue).

MeSH Terms
Amino Acid Sequence Base Sequence Catalysis Chromatography, High Pressure Liquid Cloning, Molecular DNA Modification Methylases Deoxyribonuclease EcoRI/genetics Escherichia coli/genetics Isoelectric Point Molecular Sequence Data Molecular Weight Regulatory Sequences, Nucleic Acid Rhodobacter sphaeroides/enzymology Sequence Homology, Nucleic Acid Site-Specific DNA-Methyltransferase (Adenine-Specific)/genetics,isolation & purification,metabolism Substrate Specificity
Chemicals
DNA Modification Methylases DNA modification methylase EcoRI Site-Specific DNA-Methyltransferase (Adenine-Specific) Deoxyribonuclease EcoRI
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kaszubska W
Department of Biochemistry, University of Illinois, Urbana 61801.
Aiken C
O'Connor C D
Gumport R I
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.24.10403
Published
1989-12-25
Pages
10403-25
Language
English
Region
England
NLM ID
0411011
PMCID
PMC335309
Subset
IM
Grants
NIGMS NIH HHS · GM25621 · United States
Databases
GENBANK
X16456
External Links
PubMed source DOI: 10.1093/nar/17.24.10403 PMC Full Text View journal details
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