COMPLEMENTATION OF LETHAL CONDITIONAL MUTANT YEAST BY RPL10 FROM Arabidopsis thaliana

MA LORENA FALCONE FERREYRA; MARIANELA LUCIANI D´OTTAVIO; ALEJANDRO PEZZA; PAULA CASATI

Puerto Madryn

Congreso; 46 Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2010

Sociedad Argentina de Bioquímica y Biología Vegetal

Ribosomal protein L10 (RPL10) is a key factor in joining the 40Sand 60S ribosomal subunits into a functional 80S ribosome. Inyeast, a mutation in Rpl10 demonstrated that it is essential forviability. The Arabidopsis genome contains three RPL10 genesencoding RPL10A-C. Previously, we showed that A. thalianaRPL10s are not functionally equivalent, being involved in plantdevelopment, translation and UV-B response. On the basis of thehigh degree of amino acid sequence identity between Arabidopsisand yeast RPL10s (67%); the objective of this work was to analyzethe ability of AtRPL10s to complement a yeast lethal conditionalmutant. We used the AJY2104 strain where RPL10 is expressedunder a GAL1 promoter, consequently, this strain is unviable underrepressing condition on glucose unless a functional RPL10 isepisomally provided. First, cDNAs encoding to AtRPL10A-C fromA. thaliana were cloned under control of a constitutive GDPpromoter. After transformation with these constructs,complementation was evaluated. Serial dilutions of the AtRPL10sexpressed in AJY2104 (GAL1::RPL10) showed growth ontoselective plates under repressing condition indicatingcomplementation by AtRPL10s. This result was also supported bygrowth assay in liquid medium. Future studies of polysome profileswill allow analyzing the 60S biogenesis and translation in thiscomplemented strain.