2003-01277A

FIGS1.pdf(323k)(PDF file)

Fig S1 Locations of Rad9, Rad53 and Rad24 protein binding on chromosome VI.

Locations of Rad9, Rad53 and Rad24 protein binding in the presence of 200mM HU. DMA was used for the detection of Rad53 location.

FIGS2.pdf(444k)(PDF file)

Fig S2 Interactions of checkpoint proteins with chromosome VI during normal S-phase progression.

Locations of a) Mrc1 (tagged by 3xFLAG), b) Ddc1 (tagged by 3xFLAG) and c) Ddc2 (tagged by 3xHA) after the release from -factor arrest. For Ddc1 and Ddc2, profilse of 50 min. after the release are shown.

FIGS3.pdf(528k)(PDF file)

Fig. S3 Locations of Cdc45 and replicated regions in checkpoint mutant strains treated with HU.

Locations of Cdc45 are shown together with BrdU-substituted regions in a) rad9 deletion mutant, b) tof1 deletion mutant, c) sml1 deletion mutant, and d) sml1 mec1 tel1 triple deletion mutant. All mutant strains were originated from wild type strain, SKY020 (Cdc45-3xHA GPD-TK7X). In all cases, cell cultures were prepared as in Fig.1 c) and then used for ChIP-chip.

FIGS4.pdf(1116k)(PDF file)

Fig.S4. Location of Cdc45 relative to BrdU incorporated regions in wild type and mrc1 cells treated with HU.

Wild type (a & b) and mrc1 mutant (c & d) cells were arrested in G1 phase by -factor and released in the presence of 200 mM HU at 23°C. BrdU was added to the culture 30 min. before release from G1 arrest to a final concentration of 200g/ml. Cells were withdrawn from the culture every 10 min. and analysed by ChIP-chip. Time after release from G1 arrest is indicated in each panel. BrdU incorporation was not detected until 30 min. after the release.

FIGS5.pdf(890k)

Fig.S5. Locations of various replication and checkpoint mediator proteins in wild type, mrc1 and tof1 deletion mutant strains arrested by HU.

Locations of a) Pol1 (tagged by 3xFLAG), b) Rfa1 (tagged by 3xFLAG), c) Dpb3 (tagged by 3xFLAG), d) Pol3 (tagged by 3xFLAG), e) Cdc47 (tagged by 3xHA) and f) Cdc54 (tagged by 3xHA) in wild type (upper panel) and mrc1 deletion mutant (lower panel) cells. g) Locations of Rad9 (tagged by 2xHA) in mrc1 deletion mutant. h) Locations of Tof1 (tagged by 3xFLAG, upper panel) and Mrc1 (tagged by 3xFLAG, lower panel) in mrc1 and tof1 deletion mutants, respectively. In all cases, cell cultures were prepared as described in methods.

01277AMIAME Checklist.doc (23k)(Microsoft Word 2000 document)