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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Köhrer, Caroline Bhattacharya, Arpita Mandal, Debabrata Rajbhandary, Uttam L. |
| Description | Author Affiliation: Bhattacharya A ( Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.); Köhrer C ( Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.); Mandal D ( Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.); RajBhandary UL ( Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139 bhandary@mit.edu.); |
| Abstract | 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 ß-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. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 19 |
| Volume Number | 112 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2015-05-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Archaea Genetics Codon, Terminator Haloferax Volcanii RNA, Transfer Metabolism Suppression, Genetic Codon, Nonsense Escherichia Coli Genes, Suppressor Molecular Sequence Data Novobiocin Chemistry Plasmids Promoter Regions, Genetic Serine Thymidine Tryptophan Uracil Beta-Galactosidase Research Support, N.I.H., Extramural Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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