Related Experiment Videos
A novel DNA-binding protein associated with DNA polymerase-alpha in pea stimulates polymerase activity on
J A Bryant1, D C Brice, P N Fitchett
1School of Biological Sciences, University of Exeter, Exeter EX4 4QG, UK. Department of Biological Sciences, University of Illinois, Chicago, IL 60607-7060, USA. J.A.Bryant@exeter.ac.uk
Abstract:
A 42 kDa DNA-binding protein is associated with DNA polymerase-alpha-primase in pea (Pisum sativum). In a previous publication it was shown that the protein has strong preference for ds-ss junctions in DNA, including the cohesive termini generated by restriction endonucleases. In this paper it is shown that when the DNA-binding protein is added back to polymerase-primase, the protein stimulates the activity of the polymerase. The stimulation is particularly marked when M13 DNA, primed with a single sequencing primer or primed with oligoribonucleotides by the polymerase's associated primase activity, is used as a template. The stimulation of polymerase activity is not caused by an increase in processivity. These data lead to the suggestion that the 42 kDa DNA-binding protein is a primer-recognition protein.