A novel stimulating protein of mammalian DNA polymerase α

Shonen Yoshida, Katsuyuki Tamai, Hayato Umekawa, Motoshi Suzuki, Kiyohide Kojima

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A DNA polymerase α-primase complex, which had been purified by means of immunoaffi-nity column chromatography, showed little activity in a reaction mixture composed of Tris-HCl buffer, but showed full activity in potassium phosphate buffer. It was found that potassium ion is required for the reaction by the immunoaffinity-purified enzyme. On the other hand, the DNA polymerase α purified by the orthodox biochemical method showed full activity in both buffer systems. A protein factor, which could restore the activity of immunoaffinity-purified DNA polymeraseα-primase complex in the potassium-free reaction mixture, was separated from biochemically purified DNA polymerase a. The factor, designated as factor T, was stable to heat up to 70°C, but was sensitive to trypsin. It sedimented at about 4S through a glycerol gradient. SDS-polyacrylamide gel electropho-resis revealed two polypeptide bands at 56 and 54 kDa. By immunoprecipitation, the factor T was shown to be physically associated with DNA polymerase α-primase complex. The stimulation was also observed with poly[d(A-T)], primed M13 DNA, and heat-denatured DNA.

Original languageEnglish
Pages (from-to)389-395
Number of pages7
JournalJournal of Biochemistry
Volume106
Issue number3
DOIs
Publication statusPublished - 01-01-1989

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DNA-Directed DNA Polymerase
DNA Primase
Buffers
Proteins
Potassium
Hot Temperature
Column chromatography
Tromethamine
DNA
Immunoprecipitation
Glycerol
Trypsin
Chromatography
Ions
Peptides
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Yoshida, Shonen ; Tamai, Katsuyuki ; Umekawa, Hayato ; Suzuki, Motoshi ; Kojima, Kiyohide. / A novel stimulating protein of mammalian DNA polymerase α. In: Journal of Biochemistry. 1989 ; Vol. 106, No. 3. pp. 389-395.
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A novel stimulating protein of mammalian DNA polymerase α. / Yoshida, Shonen; Tamai, Katsuyuki; Umekawa, Hayato; Suzuki, Motoshi; Kojima, Kiyohide.

In: Journal of Biochemistry, Vol. 106, No. 3, 01.01.1989, p. 389-395.

Research output: Contribution to journalArticle

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