The spatial arrangement of ORC binding modules determines the functionality of replication origins in budding yeast

Yung Tsi Bolon, Anja Katrin Bielinsky

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

In the quest to define autonomously replicating sequences (ARSs) in eukaryotic cells, an ARS consensus sequence (ACS) has emerged for budding yeast. This ACS is recognized by the replication initiator, the origin recognition complex (ORC). However, not every match to the ACS constitutes a replication origin. Here, we investigated the requirements for ORC binding to origins that carry multiple, redundant ACSs, such as ARS603. Previous studies raised the possibility that these ACSs function as individual ORC binding sites. Detailed mutational analysis of the two ACSs in ARS603 revealed that they function in concert and give rise to an initiation pattern compatible with a single bipartite ORC binding site. Consistent with this notion, deletion of one base pair between the ACS matches abolished ORC binding at ARS603. Importantly, loss of ORC binding in vitro correlated with the loss of ARS activity in vivo. Our results argue that replication origins in yeast are in general comprised of bipartite ORC binding sites that cannot function in random alignment but must conform to a configuration that permits ORC binding. These requirements help to explain why only a limited number of ACS matches in the yeast genome qualify as ORC binding sites.

Original languageEnglish (US)
Pages (from-to)5069-5080
Number of pages12
JournalNucleic acids research
Volume34
Issue number18
DOIs
StatePublished - Oct 15 2006

Fingerprint Dive into the research topics of 'The spatial arrangement of ORC binding modules determines the functionality of replication origins in budding yeast'. Together they form a unique fingerprint.

Cite this