Nucleic Acids Research, Vol 25, Issue 21 4250-4256, Copyright © 1997 by Oxford University Press
S Wiltshire, S Raychaudhuri and S Eisenberg
Although it has been demonstrated that eukaryotic cellular origins of DNA
replication may harbor stimulatory elements that bind transcription
factors, how these factors stimulate origin function is unknown. In
Saccharomyces cerevisiae , the transcription factor Abf1p stimulates origin
function of ARS121 and ARS1 . In the results presented here, an analysis of
Abf1p function has been carried out utilizing LexA(BD)- Abf1p fusion
proteins and an ARS 121 derivative harboring LexA DNA- binding sites. A
minimal region which stimulates origin function mapped to 50 amino acids
within the C-terminus of Abf1p. When tested for transcriptional activation
of a LacZ reporter gene, the same LexA(BD)- Abf1p fusion protein had
negligible transcriptional activation potential. Therefore, stimulation of
ARS 121 may occur independently of a transcriptional activation domain. It
has been previously observed that the Gal4p, Rap1p DNA-binding sites and
the LexA-Gal4p fusion protein can replace the role of Abf1p in stimulating
ARS 1 . Here we show that the stimulatory function of Abf1p at ARS 121
cannot be replaced by these alternative DNA-binding sites and the potent
chimeric transcriptional activator LexA(BD)-Gal4(AD)p . Hence, these
results strongly suggest that the Abf1p stimulation of replication may
differ for ARS 121 and ARS 1 , and imply specificity in the Abf1p/ARS 121
relationship.
ARTICLES
An Abf1p C-terminal region lacking transcriptional activation potential stimulates a yeast origin of replication
Department of Microbiology, School of Medicine, The University of Connecticut Health Center, Farmington, CT 06030, USA.
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