tailieunhanh - Báo cáo Y học: Dinucleoside polyphosphates stimulate the primer independent synthesis of poly(A) catalyzed by yeast poly(A) polymerase

Novel properties of the primer independent synthesis of poly(A), catalyzed by the yeast poly(A) polymerase are commercial enzyme fromyeast, incontrast to theenzymefromEscherichia coli, is unable to adenylate the 3¢-OH end of nucleosides, nucleotides or dinucleoside poly-phosphates (NpnN). | Eur. J. Biochem. 269 5323-5329 2002 FEBS 2002 doi Dinucleoside polyphosphates stimulate the primer independent synthesis of poly A catalyzed by yeast poly A polymerase Maria A. Giinther Sillero Anabel de Diego Hugo Osorio and Antonio Sillero Departamento de Bioquimica Instituto de Investigaciones Biomedicas Alberto Sols UAM CSIC Facultad de Medicina Madrid Spain Novel properties of the primer independent synthesis of poly A catalyzed by the yeast poly A polymerase are presented. The commercial enzyme from yeast in contrast to the enzyme from Escherichia coli is unable to adenylate the 3 -OH end of nucleosides nucleotides or dinucleoside polyphosphates NpnN . In the presence of mM ATP dinucleotides at mM activated the enzyme velocity in the following decreasing order Gp4G 100 Gp3G 82 Ap6A 61 Gp2G 52 Ap4A 51 Ap2A 41 Gp5G 36 Ap5A 27 Ap3A 20 where 100 represents a 10-fold activation in relation to a control without effector. The velocity of the enzyme towards its substrate ATP displayed sigmoidal kinetics with a Hill coefficient nH of and a Km value of mM. Dinucleoside polyphosphates did not affect the maximum velocity Vmax of the reaction but did alter its nH and Km values. In the presence of mM Gp4G or Ap4A the nH and Km values were and mM and and mM respectively. With these kinetic properties a dinucleoside polyphosphate concentration as low as 1 M may have a noticeable activating effect on the synthesis of poly A by the enzyme. These findings together with previous publications from this laboratory point to a potential relationship between dinucleoside polyphosphates and enzymes catalyzing the synthesis and or modification of DNA or RNA. Keywords Ap4A Gp4G dinucleoside polyphosphates yeast poly A polymerase. We have recently shown that Escherichia coli poly A polymerase adenylates the 3 -OH end of nucleosides nucleotides and dinucleotides of the type .

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