The centromeric regions of all Saccharomyces cerevisiae chromosomes are found in 
early replicating domains, a property conserved among centromeres in fungi and 
some higher eukaryotes. Surprisingly, little is known about the biological 
significance or the mechanism of early centromere replication; however, the 
extensive conservation suggests that it is important for chromosome maintenance. 
Do centromeres ensure their early replication by promoting early activation of 
nearby origins, or have they migrated over evolutionary time to reside in early 
replicating regions? In Candida albicans, a neocentromere contains an early 
firing origin, supporting the first hypothesis but not addressing whether the 
new origin is intrinsically early firing or whether the centromere influences 
replication time. Because the activation time of individual origins is not an 
intrinsic property of S. cerevisiae origins, but is influenced by surrounding 
sequences, we sought to test the hypothesis that centromeres influence 
replication time by moving a centromere to a late replication domain. We used a 
modified Meselson-Stahl density transfer assay to measure the kinetics of 
replication for regions of chromosome XIV in which either the functional 
centromere or a point-mutated version had been moved near origins that reside in 
a late replication region. We show that a functional centromere acts in cis over 
a distance as great as 19 kb to advance the initiation time of origins. Our 
results constitute a direct link between establishment of the kinetochore and 
the replication initiation machinery, and suggest that the proposed higher-order 
structure of the pericentric chromatin influences replication initiation.