| Title: |
Effect of the orientation and spacing of Ste12 binding sites on the expression output. |
| Authors: |
Sandrine Pinheiro; Mariona Nadal-Ribelles; Carme Solé; Vincent Vincenzetti; Yves Dusserre; Francesc Posas; Serge Pelet |
| Publication Year: |
2025 |
| Collection: |
The University of Auckland: Figshare |
| Subject Terms: |
Microbiology; Cell Biology; Genetics; Molecular Biology; Immunology; Developmental Biology; Science Policy; Infectious Diseases; Plant Biology; Biological Sciences not elsewhere classified; two haploid cells; saccharomyces cerevisiae < p; core promoter modulate; ste12 binding sites; binding sites; strong dimer; specific timing; largely encoded; large diversity; basal association |
| Description: |
A. Scheme of the two reporters present in the strains. The reference p AGA1 -dPSTR Y and a test construct, here the p SYN 3TT -dPSTR R . Both constructs encode the production of a constitutively expressed fluorescent protein and an inducible peptide containing two nuclear localization sequences (NLS) and a SynZip (SZ1 or SZ3, which can form a strong heterodimer with SZ2 and SZ4, respectively). The reference dPSTR Y is under the control of the AGA1 promoter in the yellow channel and the test dPSTR R is regulated by a synthetic promoter of interest based on a modified CYC1 promoter. B. Images of cells induced with 1µM α-factor at time 0. The nuclear enrichment of the fluorescent proteins serves as a measure of promoter activity. The scale bar represents 5 µm. C. Matrix representing the mean expression output for p CYC1 containing two PRE sites with four different orientations and spaced between 3 to 40 bp. The strength of the red color is proportional to the expression output of the promoter. Dark gray areas represent construct where fewer than 10% of cells overcome the expression threshold. Light gray squares are PRE conformations that were not measured. D. Time course of the nuclear enrichment of the dPSTR R for various distances of PRE placed in tail-to-tail orientation. The three functional conformations are plotted in blue (3 bp) light blue (13 bp) and green (23 bp). The solid lines represent the median and the shaded area, the 25- to 75-percentile of the population. Gray lines represent the median of non-functional PRE conformations. The black dashed line is the median of the control synthetic promoter without PREs inserted. E. Summary graph displaying the expression output, the speed and the fraction of responding cells for various spacings of the PRE dimer placed in the tail-to-tail orientation. The color of the marker indicates the difference in response time between the synthetic promoter and the reference p AGA1 -dPSTR Y , with fast responding promoters in red and slow ones in blue as indicated by the ... |
| Document Type: |
still image |
| Language: |
unknown |
| Relation: |
https://figshare.com/articles/figure/Effect_of_the_orientation_and_spacing_of_Ste12_binding_sites_on_the_expression_output_/29332273 |
| DOI: |
10.1371/journal.pgen.1011710.g001 |
| Availability: |
https://doi.org/10.1371/journal.pgen.1011710.g001; https://figshare.com/articles/figure/Effect_of_the_orientation_and_spacing_of_Ste12_binding_sites_on_the_expression_output_/29332273 |
| Rights: |
CC BY 4.0 |
| Accession Number: |
edsbas.1EE2E10F |
| Database: |
BASE |