Suppressors of translation initiation defect in hem12 locus of Saccharomyces cerevisiae.

• Authors: Monika Góra , Krzysztof Pluta , Anna Chełstowska , Teresa Żołądek
• Languages of publication: EN

Abstracts

EN

A system for the positive selection of transational initiation suppressors in S. cerevisiae has been developed. A mutant with an ATA initiation codon in the HEM12 gene, encoding uroporphyrinogen decarboxylase, was used to select cis- and trans-acting suppressors. These suppressors partially restore growth on nonfermentable carbon sources, such as glycerol, but still allow the accumulation of porphyrins. All extragenic suppressors are mapped to the SUI1 locus, encoding initiation factor eIF1. The effect of the hem12 mutation is also partially reversed by the known SUI3 suppressor encoding the β subunit of eIF2. In contrast, the sui2 suppressor encoding the α subunit of eIF2 does not affect the hem 12 phenotype. The intragenic suppressors are able to restore the translation of hem12 due to the generation of additional, in frame AUG codons upstream of the hem12-14 mutation. Mutational analysis of the HEM12 leader sequence was also performed to determine the role of small open reading frames (uORFs) present upstream of the HEM12 ORF. Studies on the expression of integrated hem12-1/4-lacZ fusion, devoid of all upstream ATGs, indicate a lack of regulatory effect of uORFs on HEM12 translation.

Keywords

EN

uroporphyrinogen decarboxylase; eIF2; eIF1; translation initiation; yeast

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2000
• Volume: 47
• Issue: 1
• Pages: 181-190

Dates

published

2000

received

1999-12-22

Contributors

author

Monika Góra

• Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, A. Pawińskiego 5a, 02-106 Warszawa, Poland

author

Krzysztof Pluta

• Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, A. Pawińskiego 5a, 02-106 Warszawa, Poland

author

Anna Chełstowska

• Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, A. Pawińskiego 5a, 02-106 Warszawa, Poland

author

Teresa Żołądek

• Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, A. Pawińskiego 5a, 02-106 Warszawa, Poland

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