Aphidicolin Sensitivity of Yeast and Neurospora DNA
polymerases.
Nawin Mishra (1)
(1) Dept. Biological
Sciences, University of South Carolina, Columbia, SC, USA (mishra@biol.sc.edu)
Permeabilized Yeast and Neurospora cells were incubated in presence of
[alpha-32P]dNTP and then the nature of radiolabeled products of DNA
synthesis were analysed by gel electrophoresis and quatified by
densitometry. Intermediates in the range of 50-1300 bp were found to
be synthesized which were rapidly elongated and then ligated into
longer DNA polymers. Replication inhibitors such as aphidicolin,
ara-CTP, BuPdGTP and CCA adversely affected the synthesis of DNA
pol-alpha where as carbonyldiphosphonate prevented the elongation by
DNApol-delta of the fragments generated by the action of DNA
pol-alpha. In the aphidicolin resistant mutants the synthesis of DNA
replication intermediates was not affected by phidicolin. Results of
our study, for the first time, provide insight into the role of DNA
polymerases in in vivo replication