Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Description of the genetic screening where biz1 was found. (A) Scheme of the fuz7DD allele. Two amino acid substitutions, S259D and T263D mimic the phosphorylation induced by the MAPKKK Kpp4/Ubc3. (B) Scheme of fuz7DDcrg1 allele. The endogenous fuz7 locus was exchanged by using homologous recombination with a construction carrying the mutated allele fuz7DD under the control of crg1 promoter as well as a gene conferring carboxine resistance (cbx^R). Part of the endogenous fuz7 promoter region was deleted during the insertion process. (C) Colony morphology of UMN1 strain growing in non-inducing conditions (YPD) and inducing conditions (YPA). Colonies were grown for 3 days at 28°C. Bar: 5 mm. (D) Scheme of a typical mutagenesis experiment. Liquid cultures of UMN1 grown in YPD medium to exponential phase (OD₆₀₀ around 0,6), were exposed to ultraviolet light for sufficient time to kill 80-90% of the cells, dilutions were plated directly on YPA solid medium. Petri plates were incubated for three days at 28°C and colonies with glossy appearance were isolated and saved for further analysis.
• Supplemental Figure 2 - Supplemental Figure 2. Identification of biz1, encoding a zinc finger-containing protein. (A) Colony and cell morphology of mutant fai strains. Three independent mutants are showed. Mut#2 correspond to one of the mutants complemented with a plasmid carrying kpp2. Mut#1 and Mut#3 are within the complementation process. (B) Phenotype of wild-type cells (FB1) carrying the self-replicating vector pCM54 or the pGa plasmid that harbors a DNA genomic fragment that includes the biz1 open reading frame. On solid YPD medium, FB1/pGa forms colonies with a fuzzy appearance (left panels, Bar 5 mm); in liquid culture, FB1/pGa cells show an elongated morphology (right panels, Bar 20 μm). (C) The biz1 open reading frame is highly expressed in the context of the cloning vector. Strains indicated at the top were incubated in YPD liquid medium; 10 μg of total RNA was loaded per lane. The same filter was hybridized in succession with probes for biz1 and 18s rRNA as loading control.
• Supplemental Results - Genetic screening for biz1.
• Supplemental Table 1 - Primers used in ChIP assays.