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 language | English |
|---|---|
| Pages (from-to) | 4897-4904 |
| Number of pages | 8 |
| Journal | Biochemistry |
| Volume | 8 |
| Issue number | 12 |
| State | Published - Dec 1 1969 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
Cite this
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 journal › Article
}
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.
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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 -