Absence of Replication fork associated factor CTF4 and F-box motif Encoding Gene SAF1 leads to reduction in Cell Size and Stress Tolerance Phenotype inS. cerevisiae

Abstract

14Chromosome transmission fidelity factor, Ctf4 in S. cerevisiae associates with replication fork 15 and helps in the sister chromatid cohesion. At the replication fork, Ctf4 links DNA helicase with 16 the DNA polymerase. The absence of Ctf4 invokes replication checkpoint in the cells. The Saf1 17 of S.cerevisiae interacts with Skp1 of SCF-E3 ligase though F box-motif and ubiquitinates the 18 adenine deaminase Aah1 during phase transition due to nutrient stress. The genetic interaction 19 between the CTF4 and SAF1 has not been studied. Here we report genetic interaction between 20 2 CTF4 and SAF1 which impacts the growth fitness and response to stress. The single and double 21 gene deletions of SAF1 and CTF4 were constructed in the BY4741 genetic background. The 22 strains were tested for growth on rich media and media containing stress causing agents. The 23 saf1Δctf4Δ cells with reduced cell size showed the fastest growth phenotype on YPD medium 24 when compared with the saf1Δ, ctf4Δ, and WT. The saf1Δctf4Δ cells also showed the tolerance to 25 MMS, NaCl, Glycerol, SDS, Calcofluor white, H 2 O 2 , DMSO, Benomyl, and Nocodazole when 26 compared with the saf1Δ, ctf4Δ, and WT cells. However, saf1Δctf4Δ cells showed the sensitivity 27 to HU when compared with WT and saf1Δ. Based on these observations we suggest that SAF1 28 and CTF4 interact genetically to regulate the cell size, growth and stress response. 29

Genetic interaction between F‐box motif encoding YDR131C and retrograde signaling‐related RTG1 regulates the stress response and apoptosis in Saccharomyces cerevisiae

“…For gene deletion, the homology-based one-step PCR method was adopted as mentioned in the literature. [30][31][32][33][34] The list of strains and PCR primers used in the study are mentioned in Tables 1 and 2. For cultures and maintenance of yeast strains, YPD (yeast, 1% w/v; peptone, 2% w/v; dextrose, 2% w/v; and 2% agar) and synthetic complete media (0.67% yeast nitrogen base without amino acid, 2% dextrose, and 0.2% dropout mix) were used.…”

Deletion of Autophagy gene ATG1 and F-box motif encoding gene YDR131C together leads to Synthetic Growth Defects and Flocculation behaviour in Saccharomyces cerevisiae

“…Comparative growth assessment of WT, ydr131cΔ, atg1Δ and ydr131cΔatg1Δ was carried out both on solid and liquid media as mentioned in the (S harma et al 2019) and (S alari and S alari 2017). For comparative growth assessment on solid media each strains was streaked on YPD + agar plates and incubated for 36-48 hrs at 30° C. For comparative growth kinetics of strains in the liquid media, optical density measurement at 600nm for 14 hrs was carried out.…”

“…To compare the cellular growth response of WT, ydr131cΔ, atg1Δ ,and ydr131cΔatg1Δ cells in presence of stress causing agents spot assay was performed as mentioned in ( S harma et al 2019). Briefly Wild type (BY4741) and deletion derivatives (WT, ydr131cΔ, atg1Δ and ydr131cΔatg1Δ ) were grown to log phase (OD 600 0.8-1.0) and equal number of cell were serially diluted.…”

“…To compare the cell wall status of WT, ydr131cΔ, atg1Δ , and ydr131cΔatg1Δ cells, Calcofluor white staining assay described in (P ringle 1991; P reechasuth et al 2015; S harma et al 2019) was adopted. Briefly, WT and mutant strains were grown overnight at 30ºC and next day transferred to fresh culture in 1:10 ratio.…”