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Cdc1 removes the ethanolamine phosphate of the first mannose of GPI anchors and thereby facilitates the integration of GPI proteins into the yeast cell wall.

Vazquez HM, Vionnet C, Roubaty C, Conzelmann A - Mol. Biol. Cell (2014)

Bottom Line: We find that the essential CDC1 gene can be deleted in mcd4∆ cells, which do not attach EtN-P to mannose 1 of the GPI anchor, suggesting that Cdc1 removes the EtN-P added by Mcd4.Cdc1-314(ts) mutants do not accumulate GPI proteins in the ER but have a partial secretion block later in the secretory pathway.This suggests that the presumed transglycosidases Dfg5 and Dcw1 of cdc1-314(ts) transfer GPI proteins to cell wall β1,6-glucans inefficiently.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, CH-1700 Fribourg, Switzerland.

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Genetic interactions of cdc1. (A) Fourfold dilutions of cdc1, cdc1 harboring WT CDC1, or the functionally dead cdc1D144A allele were incubated at 24 or 30°C. (B) Small areas from plates used to obtain colony sizes and Z-scores for cdc1/geneX∆ double mutants are shown. The upper three and lower three areas each show the same mutants selected either at 24, 26, or 30°C. Two double mutants (in quadruplicate) showing significant negative interaction at 24 as well as 26°C, or only at 26°C, are boxed in yellow and red, respectively, one showing positive genetic interaction is boxed in blue. The same mutants at the temperatures at which they do not get significant Z-scores are in dotted-line boxes. (C) Z-scores observed at 24 and 26°C of ∼5500 deletion strains remaining after the first filtering (see Materials and Methods) are plotted. Significant hits (Z-scores > 2.7; p values < 0.01) found only at 24°C are shown in green, hits seen only at 26°C in red, and hits found in both screens in yellow. (D–F) Negative interactions found at 24 and 26°C (D and E) and positive ones found at 30°C (F) were manually clustered into functional categories. Functional classes enriched at both 24 and 26°C are in bold. Only interactions remaining after a second filtering (see Materials and Methods) are reported.
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Figure 3: Genetic interactions of cdc1. (A) Fourfold dilutions of cdc1, cdc1 harboring WT CDC1, or the functionally dead cdc1D144A allele were incubated at 24 or 30°C. (B) Small areas from plates used to obtain colony sizes and Z-scores for cdc1/geneX∆ double mutants are shown. The upper three and lower three areas each show the same mutants selected either at 24, 26, or 30°C. Two double mutants (in quadruplicate) showing significant negative interaction at 24 as well as 26°C, or only at 26°C, are boxed in yellow and red, respectively, one showing positive genetic interaction is boxed in blue. The same mutants at the temperatures at which they do not get significant Z-scores are in dotted-line boxes. (C) Z-scores observed at 24 and 26°C of ∼5500 deletion strains remaining after the first filtering (see Materials and Methods) are plotted. Significant hits (Z-scores > 2.7; p values < 0.01) found only at 24°C are shown in green, hits seen only at 26°C in red, and hits found in both screens in yellow. (D–F) Negative interactions found at 24 and 26°C (D and E) and positive ones found at 30°C (F) were manually clustered into functional categories. Functional classes enriched at both 24 and 26°C are in bold. Only interactions remaining after a second filtering (see Materials and Methods) are reported.

Mentions: As shown in Figure 3A, cdc1* was unable to grow at 30°C; its growth was restored by WT CDC1, but not cdc1D144A, which has a point mutation in the Mn2+-binding motif (Losev et al., 2008). The CDC1* and cdc1* (cdc1-314) query strains were both crossed in quadruplicate to a library of 5850 mutants comprising deletions for each of the nonessential genes and 879 DAmP alleles of essential genes (Breslow et al., 2008).


Cdc1 removes the ethanolamine phosphate of the first mannose of GPI anchors and thereby facilitates the integration of GPI proteins into the yeast cell wall.

Vazquez HM, Vionnet C, Roubaty C, Conzelmann A - Mol. Biol. Cell (2014)

Genetic interactions of cdc1. (A) Fourfold dilutions of cdc1, cdc1 harboring WT CDC1, or the functionally dead cdc1D144A allele were incubated at 24 or 30°C. (B) Small areas from plates used to obtain colony sizes and Z-scores for cdc1/geneX∆ double mutants are shown. The upper three and lower three areas each show the same mutants selected either at 24, 26, or 30°C. Two double mutants (in quadruplicate) showing significant negative interaction at 24 as well as 26°C, or only at 26°C, are boxed in yellow and red, respectively, one showing positive genetic interaction is boxed in blue. The same mutants at the temperatures at which they do not get significant Z-scores are in dotted-line boxes. (C) Z-scores observed at 24 and 26°C of ∼5500 deletion strains remaining after the first filtering (see Materials and Methods) are plotted. Significant hits (Z-scores > 2.7; p values < 0.01) found only at 24°C are shown in green, hits seen only at 26°C in red, and hits found in both screens in yellow. (D–F) Negative interactions found at 24 and 26°C (D and E) and positive ones found at 30°C (F) were manually clustered into functional categories. Functional classes enriched at both 24 and 26°C are in bold. Only interactions remaining after a second filtering (see Materials and Methods) are reported.
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Related In: Results  -  Collection

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Figure 3: Genetic interactions of cdc1. (A) Fourfold dilutions of cdc1, cdc1 harboring WT CDC1, or the functionally dead cdc1D144A allele were incubated at 24 or 30°C. (B) Small areas from plates used to obtain colony sizes and Z-scores for cdc1/geneX∆ double mutants are shown. The upper three and lower three areas each show the same mutants selected either at 24, 26, or 30°C. Two double mutants (in quadruplicate) showing significant negative interaction at 24 as well as 26°C, or only at 26°C, are boxed in yellow and red, respectively, one showing positive genetic interaction is boxed in blue. The same mutants at the temperatures at which they do not get significant Z-scores are in dotted-line boxes. (C) Z-scores observed at 24 and 26°C of ∼5500 deletion strains remaining after the first filtering (see Materials and Methods) are plotted. Significant hits (Z-scores > 2.7; p values < 0.01) found only at 24°C are shown in green, hits seen only at 26°C in red, and hits found in both screens in yellow. (D–F) Negative interactions found at 24 and 26°C (D and E) and positive ones found at 30°C (F) were manually clustered into functional categories. Functional classes enriched at both 24 and 26°C are in bold. Only interactions remaining after a second filtering (see Materials and Methods) are reported.
Mentions: As shown in Figure 3A, cdc1* was unable to grow at 30°C; its growth was restored by WT CDC1, but not cdc1D144A, which has a point mutation in the Mn2+-binding motif (Losev et al., 2008). The CDC1* and cdc1* (cdc1-314) query strains were both crossed in quadruplicate to a library of 5850 mutants comprising deletions for each of the nonessential genes and 879 DAmP alleles of essential genes (Breslow et al., 2008).

Bottom Line: We find that the essential CDC1 gene can be deleted in mcd4∆ cells, which do not attach EtN-P to mannose 1 of the GPI anchor, suggesting that Cdc1 removes the EtN-P added by Mcd4.Cdc1-314(ts) mutants do not accumulate GPI proteins in the ER but have a partial secretion block later in the secretory pathway.This suggests that the presumed transglycosidases Dfg5 and Dcw1 of cdc1-314(ts) transfer GPI proteins to cell wall β1,6-glucans inefficiently.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, CH-1700 Fribourg, Switzerland.

Show MeSH