Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit

Motoshi Suzuki, S. Izuta, E. Savoysky, T. Sakurai, C. Simbulan, M. Tatebe, K. Kojima, S. Yoshida

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Homogeneously purified DNA polymerase α subunit-free primase was used to analyze primer RNA synthesis. On a chemically synthesized 36 mer DNA template, a part of upstream region of human c-myc gene, the primer synthesis started from a doublet of deoxythymidine (TT) in the deoxypyrimidine-rich sequence. The primase in DNA polymerase α-primase complex synthesized 21-mer reaction product, while DNA polymerase α-free primase gave the similar products, 21- and 22-mer, indicating that the site recognition was carried out by primase itself and DNA polymerase α subunit has an auxiliary role on it. Product analysis using DNA fragments carrying base substitutions further revealed that the existence of deoxypyrimidine residues around the starting sites was important for priming frequencies. Competition analysis showed that the priming was strongly competed by poly(dC), and to a much lesser extent by poly(dA). Gel-shift analysis showed that the primase could bind to the DNA template, and this complex formation was also competed by poly(dC), but not by poly(dA). These results indicate that primase subunit interacts with the starting site by binding directly with deoxypyrimidine residues.

Original languageEnglish
Pages (from-to)645-652
Number of pages8
JournalBiochemistry and Molecular Biology International
Volume29
Issue number4
Publication statusPublished - 01-01-1993

Fingerprint

DNA Primase
DNA-Directed DNA Polymerase
DNA
myc Genes
Reaction products
Thymidine
Electrophoretic Mobility Shift Assay
calf thymus DNA
Substitution reactions
Genes
Gels
Binding Sites

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Suzuki, M., Izuta, S., Savoysky, E., Sakurai, T., Simbulan, C., Tatebe, M., ... Yoshida, S. (1993). Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit. Biochemistry and Molecular Biology International, 29(4), 645-652.
Suzuki, Motoshi ; Izuta, S. ; Savoysky, E. ; Sakurai, T. ; Simbulan, C. ; Tatebe, M. ; Kojima, K. ; Yoshida, S. / Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit. In: Biochemistry and Molecular Biology International. 1993 ; Vol. 29, No. 4. pp. 645-652.
@article{79865aec36414660995518a6ef629257,
title = "Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit",
abstract = "Homogeneously purified DNA polymerase α subunit-free primase was used to analyze primer RNA synthesis. On a chemically synthesized 36 mer DNA template, a part of upstream region of human c-myc gene, the primer synthesis started from a doublet of deoxythymidine (TT) in the deoxypyrimidine-rich sequence. The primase in DNA polymerase α-primase complex synthesized 21-mer reaction product, while DNA polymerase α-free primase gave the similar products, 21- and 22-mer, indicating that the site recognition was carried out by primase itself and DNA polymerase α subunit has an auxiliary role on it. Product analysis using DNA fragments carrying base substitutions further revealed that the existence of deoxypyrimidine residues around the starting sites was important for priming frequencies. Competition analysis showed that the priming was strongly competed by poly(dC), and to a much lesser extent by poly(dA). Gel-shift analysis showed that the primase could bind to the DNA template, and this complex formation was also competed by poly(dC), but not by poly(dA). These results indicate that primase subunit interacts with the starting site by binding directly with deoxypyrimidine residues.",
author = "Motoshi Suzuki and S. Izuta and E. Savoysky and T. Sakurai and C. Simbulan and M. Tatebe and K. Kojima and S. Yoshida",
year = "1993",
month = "1",
day = "1",
language = "English",
volume = "29",
pages = "645--652",
journal = "IUBMB Life",
issn = "1521-6543",
publisher = "Wiley-Blackwell",
number = "4",

}

Suzuki, M, Izuta, S, Savoysky, E, Sakurai, T, Simbulan, C, Tatebe, M, Kojima, K & Yoshida, S 1993, 'Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit', Biochemistry and Molecular Biology International, vol. 29, no. 4, pp. 645-652.

Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit. / Suzuki, Motoshi; Izuta, S.; Savoysky, E.; Sakurai, T.; Simbulan, C.; Tatebe, M.; Kojima, K.; Yoshida, S.

In: Biochemistry and Molecular Biology International, Vol. 29, No. 4, 01.01.1993, p. 645-652.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit

AU - Suzuki, Motoshi

AU - Izuta, S.

AU - Savoysky, E.

AU - Sakurai, T.

AU - Simbulan, C.

AU - Tatebe, M.

AU - Kojima, K.

AU - Yoshida, S.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - Homogeneously purified DNA polymerase α subunit-free primase was used to analyze primer RNA synthesis. On a chemically synthesized 36 mer DNA template, a part of upstream region of human c-myc gene, the primer synthesis started from a doublet of deoxythymidine (TT) in the deoxypyrimidine-rich sequence. The primase in DNA polymerase α-primase complex synthesized 21-mer reaction product, while DNA polymerase α-free primase gave the similar products, 21- and 22-mer, indicating that the site recognition was carried out by primase itself and DNA polymerase α subunit has an auxiliary role on it. Product analysis using DNA fragments carrying base substitutions further revealed that the existence of deoxypyrimidine residues around the starting sites was important for priming frequencies. Competition analysis showed that the priming was strongly competed by poly(dC), and to a much lesser extent by poly(dA). Gel-shift analysis showed that the primase could bind to the DNA template, and this complex formation was also competed by poly(dC), but not by poly(dA). These results indicate that primase subunit interacts with the starting site by binding directly with deoxypyrimidine residues.

AB - Homogeneously purified DNA polymerase α subunit-free primase was used to analyze primer RNA synthesis. On a chemically synthesized 36 mer DNA template, a part of upstream region of human c-myc gene, the primer synthesis started from a doublet of deoxythymidine (TT) in the deoxypyrimidine-rich sequence. The primase in DNA polymerase α-primase complex synthesized 21-mer reaction product, while DNA polymerase α-free primase gave the similar products, 21- and 22-mer, indicating that the site recognition was carried out by primase itself and DNA polymerase α subunit has an auxiliary role on it. Product analysis using DNA fragments carrying base substitutions further revealed that the existence of deoxypyrimidine residues around the starting sites was important for priming frequencies. Competition analysis showed that the priming was strongly competed by poly(dC), and to a much lesser extent by poly(dA). Gel-shift analysis showed that the primase could bind to the DNA template, and this complex formation was also competed by poly(dC), but not by poly(dA). These results indicate that primase subunit interacts with the starting site by binding directly with deoxypyrimidine residues.

UR - http://www.scopus.com/inward/record.url?scp=0027312181&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027312181&partnerID=8YFLogxK

M3 - Article

C2 - 7683946

AN - SCOPUS:0027312181

VL - 29

SP - 645

EP - 652

JO - IUBMB Life

JF - IUBMB Life

SN - 1521-6543

IS - 4

ER -

Suzuki M, Izuta S, Savoysky E, Sakurai T, Simbulan C, Tatebe M et al. Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit. Biochemistry and Molecular Biology International. 1993 Jan 1;29(4):645-652.