Global mapping of translation initiation sites by TIS profiling in budding yeast
Summary: 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)...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-03-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720302379 |
| _version_ | 1818724970409230336 |
|---|---|
| author | Ina Hollerer Emily N. Powers Gloria A. Brar |
| author_facet | Ina Hollerer Emily N. Powers Gloria A. Brar |
| author_sort | Ina Hollerer |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-17T21:34:52Z |
| format | Article |
| id | doaj.art-daf0bd892ae543528367b78ea4871ad3 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-17T21:34:52Z |
| publishDate | 2021-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-daf0bd892ae543528367b78ea4871ad32022-12-21T21:31:47ZengElsevierSTAR Protocols2666-16672021-03-0121100250Global mapping of translation initiation sites by TIS profiling in budding yeastIna Hollerer0Emily N. Powers1Gloria A. Brar2Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, CA 94158, USA; Corresponding authorDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, CA 94158, USADepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, CA 94158, USA; Center for Computational Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166720302379Cell biologyGenomicsHigh-throughput screeningModel organismsMolecular biology |
| spellingShingle | Ina Hollerer Emily N. Powers Gloria A. Brar Global mapping of translation initiation sites by TIS profiling in budding yeast STAR Protocols Cell biology Genomics High-throughput screening Model organisms Molecular biology |
| title | Global mapping of translation initiation sites by TIS profiling in budding yeast |
| title_full | Global mapping of translation initiation sites by TIS profiling in budding yeast |
| title_fullStr | Global mapping of translation initiation sites by TIS profiling in budding yeast |
| title_full_unstemmed | Global mapping of translation initiation sites by TIS profiling in budding yeast |
| title_short | Global mapping of translation initiation sites by TIS profiling in budding yeast |
| title_sort | global mapping of translation initiation sites by tis profiling in budding yeast |
| topic | Cell biology Genomics High-throughput screening Model organisms Molecular biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720302379 |
| work_keys_str_mv | AT inahollerer globalmappingoftranslationinitiationsitesbytisprofilinginbuddingyeast AT emilynpowers globalmappingoftranslationinitiationsitesbytisprofilinginbuddingyeast AT gloriaabrar globalmappingoftranslationinitiationsitesbytisprofilinginbuddingyeast |