Enzymatic and chemical probing of an S1 nuclease-sensitive site upstream from the human CFTR gene

Claudia D. McDonald; Michael A. Hollingsworth; L. James Maher

doi:10.1016/0378-1119(94)90436-7

Enzymatic and chemical probing of an S1 nuclease-sensitive site upstream from the human CFTR gene

Claudia D. McDonald, Michael A. Hollingsworth, L. James Maher

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Similar purine/pyrimidine mirror repeat (PMR) DNA sequences have been identified in the 5'-flanking regions of the human cystic fibrosis transmembrane conductance regulator (hCFTR) and mucin (hMUC1) genes, and supercoiled (but not linearized) plasmids containing these promoter regions were previously shown to be sensitive to digestion by SI nuclease. The PMR element derived from the hCFTR promoter region is now sub-cloned and characterized at nucleotide resolution with respect to its reactivity toward nucleases S1 and P1, and toward the chemical probes dimethyl sulfate, chloroacetaldehyde, diethylpyrocarbonate and osmium tetroxide. These probes confirm the presence, at pH 4.5 (but not at pH 7.1), of a non-B-DNA structure. This non-B-DNA structure is distinct from H-DNA, because enzymatic and chemical probing detect single-stranded character in the absence of a stable intramolecular triple helix or extruded purine strand.

Original language	English (US)
Pages (from-to)	267-274
Number of pages	8
Journal	Gene
Volume	150
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(94)90436-7
State	Published - 1994

Keywords

DNA mirror repeats
H-DNA
PMR
cystic fibrosis
homopurine DNA
non-B-DNA

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(94)90436-7

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title = "Enzymatic and chemical probing of an S1 nuclease-sensitive site upstream from the human CFTR gene",

abstract = "Similar purine/pyrimidine mirror repeat (PMR) DNA sequences have been identified in the 5'-flanking regions of the human cystic fibrosis transmembrane conductance regulator (hCFTR) and mucin (hMUC1) genes, and supercoiled (but not linearized) plasmids containing these promoter regions were previously shown to be sensitive to digestion by SI nuclease. The PMR element derived from the hCFTR promoter region is now sub-cloned and characterized at nucleotide resolution with respect to its reactivity toward nucleases S1 and P1, and toward the chemical probes dimethyl sulfate, chloroacetaldehyde, diethylpyrocarbonate and osmium tetroxide. These probes confirm the presence, at pH 4.5 (but not at pH 7.1), of a non-B-DNA structure. This non-B-DNA structure is distinct from H-DNA, because enzymatic and chemical probing detect single-stranded character in the absence of a stable intramolecular triple helix or extruded purine strand.",

keywords = "DNA mirror repeats, H-DNA, PMR, cystic fibrosis, homopurine DNA, non-B-DNA",

author = "McDonald, {Claudia D.} and Hollingsworth, {Michael A.} and Maher, {L. James}",

note = "Funding Information: We acknowledgee xperts uggestionfsr om B. Johnston, H. Htun, S. Mirkin, B. Gold, C. Price and the technical assistanceo f J. Skoog. This work was supportedb y the following grants:f rom the National Instituteso f Health, DK44762, CA57362,G M47814 and CA36727;f rom the American Cancer Society, SIG 16; a Junior Faculty ResearchA ward (to L.J.M.), from the National Cancer Institute, 5 P30 CA36727-08; and from Abbott Laboratories,a Young Investigator{\textquoteright}sA ward (to L.J.M.).",

year = "1994",

doi = "10.1016/0378-1119(94)90436-7",

language = "English (US)",

volume = "150",

pages = "267--274",

journal = "Gene",

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publisher = "Elsevier",

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T1 - Enzymatic and chemical probing of an S1 nuclease-sensitive site upstream from the human CFTR gene

AU - McDonald, Claudia D.

AU - Hollingsworth, Michael A.

AU - Maher, L. James

N1 - Funding Information: We acknowledgee xperts uggestionfsr om B. Johnston, H. Htun, S. Mirkin, B. Gold, C. Price and the technical assistanceo f J. Skoog. This work was supportedb y the following grants:f rom the National Instituteso f Health, DK44762, CA57362,G M47814 and CA36727;f rom the American Cancer Society, SIG 16; a Junior Faculty ResearchA ward (to L.J.M.), from the National Cancer Institute, 5 P30 CA36727-08; and from Abbott Laboratories,a Young Investigator’sA ward (to L.J.M.).

PY - 1994

Y1 - 1994

N2 - Similar purine/pyrimidine mirror repeat (PMR) DNA sequences have been identified in the 5'-flanking regions of the human cystic fibrosis transmembrane conductance regulator (hCFTR) and mucin (hMUC1) genes, and supercoiled (but not linearized) plasmids containing these promoter regions were previously shown to be sensitive to digestion by SI nuclease. The PMR element derived from the hCFTR promoter region is now sub-cloned and characterized at nucleotide resolution with respect to its reactivity toward nucleases S1 and P1, and toward the chemical probes dimethyl sulfate, chloroacetaldehyde, diethylpyrocarbonate and osmium tetroxide. These probes confirm the presence, at pH 4.5 (but not at pH 7.1), of a non-B-DNA structure. This non-B-DNA structure is distinct from H-DNA, because enzymatic and chemical probing detect single-stranded character in the absence of a stable intramolecular triple helix or extruded purine strand.

AB - Similar purine/pyrimidine mirror repeat (PMR) DNA sequences have been identified in the 5'-flanking regions of the human cystic fibrosis transmembrane conductance regulator (hCFTR) and mucin (hMUC1) genes, and supercoiled (but not linearized) plasmids containing these promoter regions were previously shown to be sensitive to digestion by SI nuclease. The PMR element derived from the hCFTR promoter region is now sub-cloned and characterized at nucleotide resolution with respect to its reactivity toward nucleases S1 and P1, and toward the chemical probes dimethyl sulfate, chloroacetaldehyde, diethylpyrocarbonate and osmium tetroxide. These probes confirm the presence, at pH 4.5 (but not at pH 7.1), of a non-B-DNA structure. This non-B-DNA structure is distinct from H-DNA, because enzymatic and chemical probing detect single-stranded character in the absence of a stable intramolecular triple helix or extruded purine strand.

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KW - homopurine DNA

KW - non-B-DNA

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Enzymatic and chemical probing of an S1 nuclease-sensitive site upstream from the human CFTR gene

Abstract

Keywords

ASJC Scopus subject areas

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