abp1+ and abp2+ function downstream of SPBC2A9.02 and SPAC27D7.08c to initiate DNA replication. (A) Reduced expression levels of abp1+ and abp2+ in SPBC2A9.02Δ and SPAC27D7.08cΔ. The mRNA levels were quantified by real time PCR and those of act1+ served as an internal control (n=3). The relative level in WT was designated as arbitrary unit 1. (B) Overexpression of abp1+ and abp2+ partially rescued the growth defect of SPBC2A9.02Δ and SPAC27D7.08cΔ. pREP1-abp1+ or pREP1-abp2+ were transformed into each deletion separately. pREP1-abp1+ and pJR2-41U-abp2+ were co-transformed into SPBC2A9.02Δ or SPAC27D7.08cΔ. Transformants were harvested and 5-fold serial dilutions were spotted on plates supplemented with DNA damage reagents. Plates were photographed after 3 days of incubation at 32°C. (C) Overexpression of abp1+ or abp2+ partially relieved the G1-arrest in SPBC2A9.02Δ and SPAC27D7.08cΔ. Transformants described in Figure 4B were grown to logarithmic phase and harvested for flow cytometry analysis. Reproducible results were obtained in three independent experiments.
Pan et al. BMC Genomics 2012 13:662 doi:10.1186/1471-2164-13-662