Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I

Lisa S. Chen; Michael H. Bahr; Terry L. Sheppard

doi:10.1016/S0960-894X(03)00204-X

Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I

Lisa S. Chen, Michael H. Bahr, Terry L. Sheppard

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

5 Scopus citations

Abstract

8-Chloro-2′-deoxyadenosine (8-Cl-dAdo) was incorporated into synthetic DNA oligonucleotides to determine its effects on DNA synthesis by the 3′-5′ exonuclease-free Klenow fragment of Escherichia coli DNA Polymerase I (KF^-). Single nucleotide insertion experiments were used to determine the coding potential of 8-Cl-dAdo in a DNA template. KF^- inserted TTP opposite 8-Cl-dAdo in the template, but with decreased efficiency relative to natural deoxyadenosine. Running-start primer extensions with KF^- resulted in polymerase pausing at 8-Cl-dAdo template sites during DNA synthesis. The 2′-deoxyribonucleoside triphosphate analogue, 8-Cl-dATP, was incorporated opposite thymidine (T) approximately two-fold less efficiently than dATP.

Original language	English (US)
Pages (from-to)	1509-1512
Number of pages	4
Journal	Bioorganic and Medicinal Chemistry Letters
Volume	13
Issue number	9
DOIs	https://doi.org/10.1016/S0960-894X(03)00204-X
State	Published - May 5 2003
Externally published	Yes

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmaceutical Science
Drug Discovery
Clinical Biochemistry
Organic Chemistry

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title = "Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I",

abstract = "8-Chloro-2′-deoxyadenosine (8-Cl-dAdo) was incorporated into synthetic DNA oligonucleotides to determine its effects on DNA synthesis by the 3′-5′ exonuclease-free Klenow fragment of Escherichia coli DNA Polymerase I (KF-). Single nucleotide insertion experiments were used to determine the coding potential of 8-Cl-dAdo in a DNA template. KF- inserted TTP opposite 8-Cl-dAdo in the template, but with decreased efficiency relative to natural deoxyadenosine. Running-start primer extensions with KF- resulted in polymerase pausing at 8-Cl-dAdo template sites during DNA synthesis. The 2′-deoxyribonucleoside triphosphate analogue, 8-Cl-dATP, was incorporated opposite thymidine (T) approximately two-fold less efficiently than dATP.",

author = "Chen, {Lisa S.} and Bahr, {Michael H.} and Sheppard, {Terry L.}",

note = "Funding Information: We thank our collaborators Dr. Steven Rosen, Dr. Nancy Krett, and Dr. Varsha Gandhi for insightful discussions. We acknowledge the Keck Biophysics Facility and the Analytical Services Laboratory of Northwestern University, and the University of Illinois, Urbana-Champaign Mass Spectrometry Laboratory. This work was supported by the National Institutes of Health (RO1 CA85915-02) and the Leukemia and Lymphoma Society (6505-00).",

year = "2003",

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doi = "10.1016/S0960-894X(03)00204-X",

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T1 - Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I

AU - Chen, Lisa S.

AU - Bahr, Michael H.

AU - Sheppard, Terry L.

N1 - Funding Information: We thank our collaborators Dr. Steven Rosen, Dr. Nancy Krett, and Dr. Varsha Gandhi for insightful discussions. We acknowledge the Keck Biophysics Facility and the Analytical Services Laboratory of Northwestern University, and the University of Illinois, Urbana-Champaign Mass Spectrometry Laboratory. This work was supported by the National Institutes of Health (RO1 CA85915-02) and the Leukemia and Lymphoma Society (6505-00).

PY - 2003/5/5

Y1 - 2003/5/5

N2 - 8-Chloro-2′-deoxyadenosine (8-Cl-dAdo) was incorporated into synthetic DNA oligonucleotides to determine its effects on DNA synthesis by the 3′-5′ exonuclease-free Klenow fragment of Escherichia coli DNA Polymerase I (KF-). Single nucleotide insertion experiments were used to determine the coding potential of 8-Cl-dAdo in a DNA template. KF- inserted TTP opposite 8-Cl-dAdo in the template, but with decreased efficiency relative to natural deoxyadenosine. Running-start primer extensions with KF- resulted in polymerase pausing at 8-Cl-dAdo template sites during DNA synthesis. The 2′-deoxyribonucleoside triphosphate analogue, 8-Cl-dATP, was incorporated opposite thymidine (T) approximately two-fold less efficiently than dATP.

AB - 8-Chloro-2′-deoxyadenosine (8-Cl-dAdo) was incorporated into synthetic DNA oligonucleotides to determine its effects on DNA synthesis by the 3′-5′ exonuclease-free Klenow fragment of Escherichia coli DNA Polymerase I (KF-). Single nucleotide insertion experiments were used to determine the coding potential of 8-Cl-dAdo in a DNA template. KF- inserted TTP opposite 8-Cl-dAdo in the template, but with decreased efficiency relative to natural deoxyadenosine. Running-start primer extensions with KF- resulted in polymerase pausing at 8-Cl-dAdo template sites during DNA synthesis. The 2′-deoxyribonucleoside triphosphate analogue, 8-Cl-dATP, was incorporated opposite thymidine (T) approximately two-fold less efficiently than dATP.

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Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I

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