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Nucleic Acids Research, 2000, Vol. 28, No. 2 620-625
© 2000 Oxford University Press

The N-terminal region of DNA polymerase {delta} catalytic subunit is necessary for holoenzyme function

Sylvie Briand Schumacher, Manuel Stucki and Ulrich Hübscher*

Institute of Veterinary Biochemistry, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland

Genetic and biochemical studies have shown that DNA polymerase {delta} (Pol{delta}) is the major replicative Pol in the eukaryotic cell. Its functional form is the holoenzyme composed of Pol{delta}, proliferating cell nuclear antigen (PCNA) and replication factor C (RF-C). In this paper, we describe an N-terminal truncated form of DNA polymerase {delta} ({Delta}N Pol{delta}) from calf thymus. The {Delta}N Pol{delta} was stimulated as the full-length Pol{delta} by PCNA in a RF-C-independent Pol{delta} assay. However, when tested for holoenzyme function in a RF-C-dependent Pol{delta} assay in the presence of RF-C, ATP and replication protein A (RP-A), the {Delta}N Pol{delta} behaved differently. First, the {Delta}N Pol{delta} lacked holoenzyme functions to a great extent. Second, product size analysis and kinetic experiments showed that the holoenzyme containing {Delta}N Pol{delta} was much less efficient and synthesized DNA at a much slower rate than the holoenzyme containing full-length Pol{delta}. The present study provides the first evidence that the N-terminal part of the large subunit of Pol{delta} is involved in holo­enzyme function.

* To whom correspondence should be addressed. Tel: +41 1 635 5472; Fax: +41 1 635 6840; Email: hubscher@vetbio.unizh.ch


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