Nonsense suppression in archaea
Bacterial strains carrying nonsense suppressor tRNA genes played a crucial role in early work on bacterial and bacterial viral genetics. In eukaryotes as well, suppressor tRNAs have played important roles in the genetic analysis of yeast and worms. Surprisingly, little is known about genetic suppres...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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National Academy of Sciences (U.S.)
2016
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| Online Access: | http://hdl.handle.net/1721.1/100740 https://orcid.org/0000-0002-4530-5647 |
| _version_ | 1826198698356375552 |
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| author | Bhattacharya, Arpita Mandal, Debabrata Koehrer, Caroline Rajbhandary, Uttam L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Bhattacharya, Arpita Mandal, Debabrata Koehrer, Caroline Rajbhandary, Uttam L |
| author_sort | Bhattacharya, Arpita |
| collection | MIT |
| description | Bacterial strains carrying nonsense suppressor tRNA genes played a crucial role in early work on bacterial and bacterial viral genetics. In eukaryotes as well, suppressor tRNAs have played important roles in the genetic analysis of yeast and worms. Surprisingly, little is known about genetic suppression in archaea, and there has been no characterization of suppressor tRNAs or identification of nonsense mutations in any of the archaeal genes. Here, we show, using the β-gal gene as a reporter, that amber, ochre, and opal suppressors derived from the serine and tyrosine tRNAs of the archaeon Haloferax volcanii are active in suppression of their corresponding stop codons. Using a promoter for tRNA expression regulated by tryptophan, we also show inducible and regulatable suppression of all three stop codons in H. volcanii. Additionally, transformation of a ΔpyrE2 H. volcanii strain with plasmids carrying the genes for a pyrE2 amber mutant and the serine amber suppressor tRNA yielded transformants that grow on agar plates lacking uracil. Thus, an auxotrophic amber mutation in the pyrE2 gene can be complemented by expression of the amber suppressor tRNA. These results pave the way for generating archaeal strains carrying inducible suppressor tRNA genes on the chromosome and their use in archaeal and archaeviral genetics. We also provide possible explanations for why suppressor tRNAs have not been identified in archaea. |
| first_indexed | 2024-09-23T11:08:44Z |
| format | Article |
| id | mit-1721.1/100740 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:08:44Z |
| publishDate | 2016 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1007402022-09-27T17:28:24Z Nonsense suppression in archaea Bhattacharya, Arpita Mandal, Debabrata Koehrer, Caroline Rajbhandary, Uttam L Massachusetts Institute of Technology. Department of Biology Bhattacharya, Arpita Koehrer, Caroline Mandal, Debabrata Rajbhandary, Uttam L. Bacterial strains carrying nonsense suppressor tRNA genes played a crucial role in early work on bacterial and bacterial viral genetics. In eukaryotes as well, suppressor tRNAs have played important roles in the genetic analysis of yeast and worms. Surprisingly, little is known about genetic suppression in archaea, and there has been no characterization of suppressor tRNAs or identification of nonsense mutations in any of the archaeal genes. Here, we show, using the β-gal gene as a reporter, that amber, ochre, and opal suppressors derived from the serine and tyrosine tRNAs of the archaeon Haloferax volcanii are active in suppression of their corresponding stop codons. Using a promoter for tRNA expression regulated by tryptophan, we also show inducible and regulatable suppression of all three stop codons in H. volcanii. Additionally, transformation of a ΔpyrE2 H. volcanii strain with plasmids carrying the genes for a pyrE2 amber mutant and the serine amber suppressor tRNA yielded transformants that grow on agar plates lacking uracil. Thus, an auxotrophic amber mutation in the pyrE2 gene can be complemented by expression of the amber suppressor tRNA. These results pave the way for generating archaeal strains carrying inducible suppressor tRNA genes on the chromosome and their use in archaeal and archaeviral genetics. We also provide possible explanations for why suppressor tRNAs have not been identified in archaea. National Institutes of Health (U.S.) (Grant GM17151) 2016-01-07T01:52:08Z 2016-01-07T01:52:08Z 2015-05 2015-01 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/100740 Bhattacharya, Arpita, Caroline Kohrer, Debabrata Mandal, and Uttam L. RajBhandary. “Nonsense Suppression in Archaea.” Proc Natl Acad Sci USA 112, no. 19 (April 27, 2015): 6015–6020. https://orcid.org/0000-0002-4530-5647 en_US http://dx.doi.org/10.1073/pnas.1501558112 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) American Meteorological Society |
| spellingShingle | Bhattacharya, Arpita Mandal, Debabrata Koehrer, Caroline Rajbhandary, Uttam L Nonsense suppression in archaea |
| title | Nonsense suppression in archaea |
| title_full | Nonsense suppression in archaea |
| title_fullStr | Nonsense suppression in archaea |
| title_full_unstemmed | Nonsense suppression in archaea |
| title_short | Nonsense suppression in archaea |
| title_sort | nonsense suppression in archaea |
| url | http://hdl.handle.net/1721.1/100740 https://orcid.org/0000-0002-4530-5647 |
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