Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases.

• 保川, 清

• Konishi, Atsushi

• Shinomura, Mayu

• Nagaoka, Eriko

• Fujiwara, Shinsuke

抄録

Some bacterial thermostable, wild-type or genetically engineered family A DNA polymerases have reverse transcriptase activity. However, difference in reverse transcriptase activities of family A DNA polymerases and retroviral reverse transcriptases (RTs) is unclear. In this study, comparative kinetic analysis was performed for the reverse transcriptase activities of the wild-type enzyme of family A DNA polymerase (M1pol(WT)) from Thermus thermophilus M1 and the variant enzyme of family A DNA polymerase (K4pol(L329A)), in which the mutation of Leu329→Ala is undertaken, from Thermotoga petrophila K4. In the incorporation of dTTP into poly(rA)-p(dT)(45), the reaction rates of K4pol(L329A) and M1pol(WT) exhibited a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)(45) concentrations. In contrast, the reaction rates of Moloney murine leukemia virus (MMLV) RT exhibited saturated profiles for both dTTP and poly(rA)-p(dT)(45) concentrations. This suggests that high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases.

収録刊行物

• Biochemical and biophysical research communications

  Biochemical and biophysical research communications 427 (3), 654-658, 2012-10-26

  Elsevier Inc.

キーワード

• Family A DNA polymerase
• Moloney murine leukemia virus
• Reverse transcriptase
• Template-primer
• Thermotoga petrophila
• Thermusthermophilus

詳細情報

• CRID: 1050564285695855232
• NII論文ID: 120004945040
• NII書誌ID: AA11542044
• ISSN: 0006291X
• HANDLE: 2433/161657
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • CiNii Articles
  • KAKEN