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Nucleic Acids Research, 1991, Vol. 19, No. 25 7081-7088
© 1991

Foreword

T-antigen binding to site I facilities initiation of SV40 DNA replication but does not affect bidirectionality

Zong-Sheng Guo+, Uwe Heine§ and L. Depamphilis*

Department of Cell and Developmental Biology, Roche Institute of Molecular Biology, Roche Research Center Nutley, NJ 07110, USA

*To whom correspondence should be addressed

SV40 origin auxiliary sequence 1 (aux–1) encompasses T-antlgen (T-ag) binding site I and facilitates origin core (on-core) activity in whole cells or cell extracts. Aux–1 activity depended completely upon its sequence, orientation and spacing relative to orf-core. Aux–1 activity was lost either by Inserting 10 base pairs between aux–1 and on-core or by placing either orientation of aux–1 on the opposite side of ori-core. Reversing the orientation of aux-1 in Its normal position actually inhibited replication. Easily unwound DNA sequences that stimulate yeast or E. coli origins of replication could not replace aux–1. Aux–1 did not affect bidirectional replication. Replication remained bidirectional even when aux–1 was inactivated, and deletion of aux–1 did not affect selection of RNA-primed DNA synthesis initiation sites in the origin region: the transition from discontinuous to continuous DNA synthesis that marks the origin of bidirectional replication occurred at the same nucleotlde locations in both wild-type and aux–1 deleted origins. These results support a model for initiation of SV40 DNA replication in which T-ag binding to aux–1 (T-ag binding site I) facilitates the efficiency with which T-ag initiates replication at on-core (T-ag binding site II) without affecting the mechanism by which initiation of DNA replication occurs.

+Present address: Howard Hughes Medical Institute, New York University Medical Center, New York, NY 1001

§Present address: Roche Biomedical Laboratories, 1447 York Court, Burlington, NC 27215, USA


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