A comprehensive tRNA deletion library unravels the genetic architecture of the tRNA pool.
Deciphering the architecture of the tRNA pool is a prime challenge in translation research, as tRNAs govern the efficiency and accuracy of the process. Towards this challenge, we created a systematic tRNA deletion library in Saccharomyces cerevisiae, aimed at dissecting the specific contribution of each tRNA gene to the tRNA pool and to the cell's fitness. By harnessing this resource, we observed that the majority of tRNA deletions show no appreciable phenotype in rich medium, yet under more challenging conditions, additional phenotypes were observed. Robustness to tRNA gene deletion was often facilitated through extensive backup compensation within and between tRNA families. Interestingly, we found that within tRNA families, genes carrying identical anti-codons can contribute differently to the cellular fitness, suggesting the importance of the genomic surrounding to tRNA expression. Characterization of the transcriptome response to deletions of tRNA genes exposed two disparate patterns: in single-copy families, deletions elicited a stress response; in deletions of genes from multi-copy families, expression of the translation machinery increased. Our results uncover the complex architecture of the tRNA pool and pave the way towards complete understanding of their role in cell physiology.
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Zohar
Z
Bloom-Ackermann
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Sivan
S
Navon
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Hila
H
Gingold
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Ruth
R
Towers
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Yitzhak
Y
Pilpel
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Orna
O
Dahan
D003062:Codon / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D056084:Genetic Fitness; D012343:RNA, Transfer / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D017384:Sequence Deletion; D013312:Stress, Physiological / Q000235:genetics*; D059467:Transcriptome
D003062:Codon; D012343:RNA, Transfer