The accessibility of DNA to dimethylsulfate in complexes with recA protein

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

recA protein coats DNA co-operatively to form filaments approximately 100 A thick, which in the presence of ATP, and more stably so in the presence of the non-hydrolyzable analog ATP gamma S, have a helical appearance with a deep cleft in the protein coat. This protein helix follows the DNA helix, to which it imparts a new helicity of 18.5 bp per turn of 97 A pitch. Here we test the accessibility of the DNA in the complex to modification by dimethylsulfate, and find that the complexed DNA is approximately 2-fold more reactive on the major groove side than it was in B-DNA (methylation of guanine N7), while it is protected approximately 2-fold on the minor groove side (methylation of adenine N3), suggesting that the protein coats the DNA along the minor groove. Furthermore, N3 of cytosine, a residue involved in base pairing, is found exposed in complexes with single strands as it is in naked single-stranded DNA, while it remains inaccessible in complexes with double strands, suggesting that the latter is not melted at this stage of the strand exchange reaction.
Original languageEnglish
Pages (from-to)2493-2498
Number of pages6
JournalEMBO Journal
Volume6
Issue number8
Publication statusPublished - 1 Aug 1987

Fingerprint

Rec A Recombinases
DNA
Methylation
Capsid Proteins
B-Form DNA
Single-Stranded DNA
Cytosine
Guanine
Adenine
Base Pairing
Adenosine Triphosphate
dimethyl sulfate
Proteins

Keywords

  • Alkylating Agents
  • Base Sequence
  • DNA, Bacterial
  • Methylation
  • Microscopy, Electron
  • Plasmids
  • Rec A Recombinases
  • Sulfuric Acid Esters
  • Sulfuric Acids

Cite this

The accessibility of DNA to dimethylsulfate in complexes with recA protein. / Di Capua, E; Muller, Berndt Marino.

In: EMBO Journal, Vol. 6, No. 8, 01.08.1987, p. 2493-2498.

Research output: Contribution to journalArticle

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abstract = "recA protein coats DNA co-operatively to form filaments approximately 100 A thick, which in the presence of ATP, and more stably so in the presence of the non-hydrolyzable analog ATP gamma S, have a helical appearance with a deep cleft in the protein coat. This protein helix follows the DNA helix, to which it imparts a new helicity of 18.5 bp per turn of 97 A pitch. Here we test the accessibility of the DNA in the complex to modification by dimethylsulfate, and find that the complexed DNA is approximately 2-fold more reactive on the major groove side than it was in B-DNA (methylation of guanine N7), while it is protected approximately 2-fold on the minor groove side (methylation of adenine N3), suggesting that the protein coats the DNA along the minor groove. Furthermore, N3 of cytosine, a residue involved in base pairing, is found exposed in complexes with single strands as it is in naked single-stranded DNA, while it remains inaccessible in complexes with double strands, suggesting that the latter is not melted at this stage of the strand exchange reaction.",
keywords = "Alkylating Agents, Base Sequence, DNA, Bacterial, Methylation, Microscopy, Electron, Plasmids, Rec A Recombinases, Sulfuric Acid Esters, Sulfuric Acids",
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T1 - The accessibility of DNA to dimethylsulfate in complexes with recA protein

AU - Di Capua, E

AU - Muller, Berndt Marino

PY - 1987/8/1

Y1 - 1987/8/1

N2 - recA protein coats DNA co-operatively to form filaments approximately 100 A thick, which in the presence of ATP, and more stably so in the presence of the non-hydrolyzable analog ATP gamma S, have a helical appearance with a deep cleft in the protein coat. This protein helix follows the DNA helix, to which it imparts a new helicity of 18.5 bp per turn of 97 A pitch. Here we test the accessibility of the DNA in the complex to modification by dimethylsulfate, and find that the complexed DNA is approximately 2-fold more reactive on the major groove side than it was in B-DNA (methylation of guanine N7), while it is protected approximately 2-fold on the minor groove side (methylation of adenine N3), suggesting that the protein coats the DNA along the minor groove. Furthermore, N3 of cytosine, a residue involved in base pairing, is found exposed in complexes with single strands as it is in naked single-stranded DNA, while it remains inaccessible in complexes with double strands, suggesting that the latter is not melted at this stage of the strand exchange reaction.

AB - recA protein coats DNA co-operatively to form filaments approximately 100 A thick, which in the presence of ATP, and more stably so in the presence of the non-hydrolyzable analog ATP gamma S, have a helical appearance with a deep cleft in the protein coat. This protein helix follows the DNA helix, to which it imparts a new helicity of 18.5 bp per turn of 97 A pitch. Here we test the accessibility of the DNA in the complex to modification by dimethylsulfate, and find that the complexed DNA is approximately 2-fold more reactive on the major groove side than it was in B-DNA (methylation of guanine N7), while it is protected approximately 2-fold on the minor groove side (methylation of adenine N3), suggesting that the protein coats the DNA along the minor groove. Furthermore, N3 of cytosine, a residue involved in base pairing, is found exposed in complexes with single strands as it is in naked single-stranded DNA, while it remains inaccessible in complexes with double strands, suggesting that the latter is not melted at this stage of the strand exchange reaction.

KW - Alkylating Agents

KW - Base Sequence

KW - DNA, Bacterial

KW - Methylation

KW - Microscopy, Electron

KW - Plasmids

KW - Rec A Recombinases

KW - Sulfuric Acid Esters

KW - Sulfuric Acids

M3 - Article

VL - 6

SP - 2493

EP - 2498

JO - EMBO Journal

JF - EMBO Journal

SN - 0261-4189

IS - 8

ER -