Caulobacter crescentus Cell Cycle-Regulated DNA Methyltransferase Uses a Novel Mechanism for Substrate Recognition

Caulobacter crescentus relies on DNA methylation by the cell cycle-regulated methyltransferase (CcrM) in addition to key transcription factors to control the cell cycle and direct cellular differentiation. CcrM is shown here to efficiently methylate its cognate recognition site 5′-GANTC-3′ in single...

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Bibliographic Details
Published in:Biochemistry (Easton) 2017-08, Vol.56 (30), p.3913-3922
Main Authors: Woodcock, Clayton B, Yakubov, Aziz B, Reich, Norbert O
Format: Article
Language:English
Subjects:
Caulobacter crescentus - cytology
Caulobacter crescentus - enzymology
Cell Cycle
Coenzymes - metabolism
DNA - chemistry
DNA - metabolism
DNA Methylation
DNA, Single-Stranded - chemistry
DNA, Single-Stranded - metabolism
Electrophoretic Mobility Shift Assay
Fluoresceins - analysis
Fluorescent Dyes - analysis
Kinetics
Nucleotide Motifs
Recombinant Fusion Proteins - metabolism
Recombinant Proteins - metabolism
RNA - chemistry
RNA - metabolism
S-Adenosylmethionine - metabolism
Site-Specific DNA-Methyltransferase (Adenine-Specific) - genetics
Site-Specific DNA-Methyltransferase (Adenine-Specific) - metabolism
Substrate Specificity
Thermodynamics
Tritium
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Summary:Caulobacter crescentus relies on DNA methylation by the cell cycle-regulated methyltransferase (CcrM) in addition to key transcription factors to control the cell cycle and direct cellular differentiation. CcrM is shown here to efficiently methylate its cognate recognition site 5′-GANTC-3′ in single-stranded and hemimethylated double-stranded DNA. We report the K m, k cat, k methylation, and K d for single-stranded and hemimethylated substrates, revealing discrimination of 107-fold for noncognate sequences. The enzyme also shows a similar discrimination against single-stranded RNA. Two independent assays clearly show that CcrM is highly processive with single-stranded and hemimethylated DNA. Collectively, the data provide evidence that CcrM and other DNA-modifying enzymes may use a new mechanism to recognize DNA in a key epigenetic process.
ISSN:0006-2960
1520-4995
DOI:10.1021/acs.biochem.7b00378