Additional file 9 of TORC1 regulates the transcriptional response to glucose and developmental cycle via the Tap42-Sit4-Rrd1/2 pathway in Saccharomyces cerevisiae
Creators: name: Alfatah, Mohammad
name: Wong, Jin Huei
name: Vidhya Gomathi Krishnan
name: Lee, Yong Cheow
name: Sin, Quan Feng
name: Goh, Corinna Jie Hui
name: Kiat Whye Kong
name: Lee, Wei Ting
name: Lewis, Jacqueline
name: Hoon, Shawn
name: Arumugam, Prakash
Publisher: name: figshare
Year: 2021
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: 69999 Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences

description: Additional file 9: Fig. S14. Overlap of glucose-responsive genes with TORC1 target genes. a Comparison of our TORC1 target list identified during spore germination with those reported in a Micro-array-based study (Shamji et al.) and an RNA-Seq study (Gowans et al.) [ both performed with vegetative cells. b Comparison of the list of glucose-responsive genes (Wang et al.) with TORC1 target lists identified by our RNA-Seq analysis and an independent RNA-Seq study (Gowans et al.). Fig. S15. TORC1 regulates the glucose-responsive genes during spore germination. Spores were transferred to YPD medium with either DMSO or rapamycin (2 μM). Aliquots of yeast cells taken at the indicated time points (0 h, 0.5 h, 1 h, and 2 h) from the two cultures were used for preparing RNA. Expression of 7 TGC genes (GFD2, GPG1, UGA1, RME1, CIT1, CRC1 and DHR2) was assayed by Real-Time qRT-PCR analysis. Levels of transcripts were normalized with respect to actin mRNA. Data are presented as means ± standard deviation (n = 2 technical replicates). Fig. S16. TORC1 is required for spore germination. a Fourteen asci resulting from sporulation of wild type or fpr1Δ or TOR1-1 diploid cells were dissected on YPD + agar plates containing 1.5 μM rapamycin. Fourteen asci from wild type diploid cells were also dissected on a YPD agar plate without rapamycin. Percentage of budded cells was calculated by examining the spore morphology under the dissection microscope after 6 hours following dissection and is indicated in the plot. b Images of the agar plates described above following incubation at 30 °C for 2 days are presented. Fig. S17. TORC1 is required for expression of cell cycle genes during spore germination. Spores were transferred to YPD medium with either DMSO or rapamycin (2 μM). Aliquots of yeast cells taken at the indicated time points (0, 4, 6 and 7 h) from the two cultures were used for preparing RNA. Expression of cell cycle genes CLN1, CLN2, CLB6, EGT2, PCL1, PCL9, CHS2 and CTS1 was assayed by Real-Time qPCR analysis. Levels of transcripts were normalized with respect to actin mRNA. Data are presented as means ± standard deviation (n = 2 replicates). Fig. S18. TORC1’s role in spore germination is mediated via inhibition of Rrd1. Wild type and rrd1Δ spores were transferred into YPD medium in the presence of either DMSO or rapamycin (2 μM). Progress of spore germination was assayed by scoring fraction of cells with different morphologies at the indicated time points for up to 8 hours following transfer into YPD medium.