Genetic interactions of a putative Arabidopsis thaliana ubiquitin-ligase with components of the Saccharomyces cerevisiae ubiquitination machinery.
The feasibility of using the Saccharomyces cerevisiae genetic tools to get insights into the function of a plant-specific ubiquitin-ligase was examined. ATL2 is a potential ubiquitin-ligase of the RING-H2 type that was originally isolated as a conditionally toxic Arabidopsis cDNA when overexpressed in yeast. ATL2 is a member of an Arabidopsis family that comprises 80 proteins. After testing cDNAs from 25 ATL members for toxicity we found that in addition to ATL2 only ATL63 was toxic, suggesting specific interactions of each one of these two ATLs in yeast. We seek to identify suppressors of the ATL2 toxicity in yeast and we found that toxicity was suppressed by knock-out mutations on different components of the ubiquitination pathway. Suppression was achieved in four deubiquitinating enzyme mutants and in one ubiquitin-conjugating enzyme mutant. A model is proposed in which Ubc4 and ATL2 act together to target for degradation one or more essential yeast proteins, Doa4/Ubp4, Ubp6 and Ubp14 have a role in disassembling ubiquitin chains on the target proteins and Ubp15 protects ATL2 from auto-ubiquitination. We presuppose that our approach can be further utilized to analyze the function of this distinctive class of ubiquitin-ligases in yeast as well as in Arabidopsis.
Aguilar-Henonin
Laura
L
Departamento de Ingeniería Genética de Plantas, Centro de Investigación y de Estudios Avanzados, Campus Guanajuato., Apartado Postal 629, Gto, 36500, Mexico.
Bravo
Jaime
J
Guzmán
Plinio
P
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D008954:Models, Biological; D005810:Multigene Family; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D013489:Suppression, Genetic; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism; D014452:Ubiquitins / Q000378:metabolism*
C102152:ATL2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D014452:Ubiquitins; C501306:Ubc4 protein, S cerevisiae; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D010450:Endopeptidases; C109028:UBP6 protein, S cerevisiae