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Global mapping of translation initiation sites by TIS profiling in budding yeast.

ABSTRACT: Translation initiation site (TIS) profiling allows for the genome-wide identification of TISs in vivo by exclusively capturing mRNA fragments within ribosomes that have just completed translation initiation. It leverages translation inhibitors, such as harringtonine and lactimidomycin (LTM), that preferentially capture ribosomes at start codon positions, protecting TIS-derived mRNA fragments from nuclease digestion. Here, we describe a step-by-step protocol for TIS profiling in LTM-treated budding yeast that we developed to identify TISs and open reading frames in vegetative and meiotic cells. For complete details on the use and execution of this protocol, please refer to Eisenberg et al. (2020).

SUBMITTER: Hollerer I

PROVIDER: S-EPMC7797918 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Global mapping of translation initiation sites by TIS profiling in budding yeast.

Hollerer Ina I Powers Emily N EN Brar Gloria A GA

STAR protocols 20201230 1

Translation initiation site (TIS) profiling allows for the genome-wide identification of TISs <i>in vivo</i> by exclusively capturing mRNA fragments within ribosomes that have just completed translation initiation. It leverages translation inhibitors, such as harringtonine and lactimidomycin (LTM), that preferentially capture ribosomes at start codon positions, protecting TIS-derived mRNA fragments from nuclease digestion. Here, we describe a step-by-step protocol for TIS profiling in LTM-treate ...[more]

PMID: 33458709

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