Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide

Maurice R. Atkinson, Murray P Deutscher, Arthur Kornberg, Alan F. Russell, J. G. Moffatt

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

2′,3′-Dideoxyribonucleoside triphosphates are analogs of the natural 2′-deoxyribonucleotide substrates of deoxyribonucleic acid polymerase but lack the 3′-hydroxyl group required for deoxyribonucleic acid chain growth. Attachment of a dideoxynucleotide blocks deoxyribonucleic acid synthesis and inhibits related reactions (pyrophosphorolysis, pyrophosphate exchange, and hydrolysis) which occur at the primer site of deoxyribonucleic acid polymerase from Escherichia coli. Attachment of a chain-terminating dideoxythymidylate group to deoxyribonucleic acid and to oligo-and polydeoxynucleotide chains is approximately a thousand times slower than that of deoxyribothymidylate. Hydrolysis and pyrophosphate exchange are inhibited to a similar extent. The requirement for a 3′-hydroxyl group for optimal rates of these reactions is discussed in terms of a model for deoxyribonucleic acid polymerase action. In the presence of an excess of polymerase, the extent of incorporation of dideoxythymidylate residues at the 3′ terminus of poly d(A-T) and of deoxyribonucleic acid chains is proportional to polynucleotide concentration, and thus permits a determination of available primer sites.

Original languageEnglish
Pages (from-to)4897-4904
Number of pages8
JournalBiochemistry
Volume8
Issue number12
StatePublished - Dec 1 1969
Externally publishedYes

Fingerprint

DNA
Growth
Hydroxyl Radical
Hydrolysis
Dideoxynucleotides
Dideoxynucleosides
Deoxyribonucleotides
Polynucleotides
Escherichia coli
Ion exchange
Substrates
diphosphoric acid

ASJC Scopus subject areas

  • Biochemistry

Cite this

Atkinson, M. R., Deutscher, M. P., Kornberg, A., Russell, A. F., & Moffatt, J. G. (1969). Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide. Biochemistry, 8(12), 4897-4904.

Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide. / Atkinson, Maurice R.; Deutscher, Murray P; Kornberg, Arthur; Russell, Alan F.; Moffatt, J. G.

In: Biochemistry, Vol. 8, No. 12, 01.12.1969, p. 4897-4904.

Research output: Contribution to journalArticle

Atkinson, MR, Deutscher, MP, Kornberg, A, Russell, AF & Moffatt, JG 1969, 'Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide', Biochemistry, vol. 8, no. 12, pp. 4897-4904.
Atkinson MR, Deutscher MP, Kornberg A, Russell AF, Moffatt JG. Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide. Biochemistry. 1969 Dec 1;8(12):4897-4904.
Atkinson, Maurice R. ; Deutscher, Murray P ; Kornberg, Arthur ; Russell, Alan F. ; Moffatt, J. G. / Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide. In: Biochemistry. 1969 ; Vol. 8, No. 12. pp. 4897-4904.
@article{16d09036109543c28a8e262dbe78b420,
title = "Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide",
abstract = "2′,3′-Dideoxyribonucleoside triphosphates are analogs of the natural 2′-deoxyribonucleotide substrates of deoxyribonucleic acid polymerase but lack the 3′-hydroxyl group required for deoxyribonucleic acid chain growth. Attachment of a dideoxynucleotide blocks deoxyribonucleic acid synthesis and inhibits related reactions (pyrophosphorolysis, pyrophosphate exchange, and hydrolysis) which occur at the primer site of deoxyribonucleic acid polymerase from Escherichia coli. Attachment of a chain-terminating dideoxythymidylate group to deoxyribonucleic acid and to oligo-and polydeoxynucleotide chains is approximately a thousand times slower than that of deoxyribothymidylate. Hydrolysis and pyrophosphate exchange are inhibited to a similar extent. The requirement for a 3′-hydroxyl group for optimal rates of these reactions is discussed in terms of a model for deoxyribonucleic acid polymerase action. In the presence of an excess of polymerase, the extent of incorporation of dideoxythymidylate residues at the 3′ terminus of poly d(A-T) and of deoxyribonucleic acid chains is proportional to polynucleotide concentration, and thus permits a determination of available primer sites.",
author = "Atkinson, {Maurice R.} and Deutscher, {Murray P} and Arthur Kornberg and Russell, {Alan F.} and Moffatt, {J. G.}",
year = "1969",
month = "12",
day = "1",
language = "English",
volume = "8",
pages = "4897--4904",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "12",

}

TY - JOUR

T1 - Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2′,3′-dideoxyribonucleotide

AU - Atkinson, Maurice R.

AU - Deutscher, Murray P

AU - Kornberg, Arthur

AU - Russell, Alan F.

AU - Moffatt, J. G.

PY - 1969/12/1

Y1 - 1969/12/1

N2 - 2′,3′-Dideoxyribonucleoside triphosphates are analogs of the natural 2′-deoxyribonucleotide substrates of deoxyribonucleic acid polymerase but lack the 3′-hydroxyl group required for deoxyribonucleic acid chain growth. Attachment of a dideoxynucleotide blocks deoxyribonucleic acid synthesis and inhibits related reactions (pyrophosphorolysis, pyrophosphate exchange, and hydrolysis) which occur at the primer site of deoxyribonucleic acid polymerase from Escherichia coli. Attachment of a chain-terminating dideoxythymidylate group to deoxyribonucleic acid and to oligo-and polydeoxynucleotide chains is approximately a thousand times slower than that of deoxyribothymidylate. Hydrolysis and pyrophosphate exchange are inhibited to a similar extent. The requirement for a 3′-hydroxyl group for optimal rates of these reactions is discussed in terms of a model for deoxyribonucleic acid polymerase action. In the presence of an excess of polymerase, the extent of incorporation of dideoxythymidylate residues at the 3′ terminus of poly d(A-T) and of deoxyribonucleic acid chains is proportional to polynucleotide concentration, and thus permits a determination of available primer sites.

AB - 2′,3′-Dideoxyribonucleoside triphosphates are analogs of the natural 2′-deoxyribonucleotide substrates of deoxyribonucleic acid polymerase but lack the 3′-hydroxyl group required for deoxyribonucleic acid chain growth. Attachment of a dideoxynucleotide blocks deoxyribonucleic acid synthesis and inhibits related reactions (pyrophosphorolysis, pyrophosphate exchange, and hydrolysis) which occur at the primer site of deoxyribonucleic acid polymerase from Escherichia coli. Attachment of a chain-terminating dideoxythymidylate group to deoxyribonucleic acid and to oligo-and polydeoxynucleotide chains is approximately a thousand times slower than that of deoxyribothymidylate. Hydrolysis and pyrophosphate exchange are inhibited to a similar extent. The requirement for a 3′-hydroxyl group for optimal rates of these reactions is discussed in terms of a model for deoxyribonucleic acid polymerase action. In the presence of an excess of polymerase, the extent of incorporation of dideoxythymidylate residues at the 3′ terminus of poly d(A-T) and of deoxyribonucleic acid chains is proportional to polynucleotide concentration, and thus permits a determination of available primer sites.

UR - http://www.scopus.com/inward/record.url?scp=0014619339&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0014619339&partnerID=8YFLogxK

M3 - Article

C2 - 4312461

AN - SCOPUS:0014619339

VL - 8

SP - 4897

EP - 4904

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 12

ER -