dc.contributor.advisor | Rodnina, Marina Prof. Dr. | de |
dc.contributor.author | Kotini, Suresh Babu | de |
dc.date.accessioned | 2012-06-27T18:35:38Z | de |
dc.date.accessioned | 2013-01-18T14:23:30Z | de |
dc.date.available | 2013-01-30T23:51:09Z | de |
dc.date.issued | 2012-06-27 | de |
dc.identifier.uri | http://hdl.handle.net/11858/00-1735-0000-000D-F0BB-9 | de |
dc.identifier.uri | http://dx.doi.org/10.53846/goediss-3190 | |
dc.format.mimetype | application/pdf | de |
dc.language.iso | eng | de |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/3.0/ | de |
dc.title | Molecular mechanism of selenocysteine incorporation in bacterial translation | de |
dc.type | doctoralThesis | de |
dc.title.translated | English | de |
dc.contributor.referee | Rodnina, Marina Prof. Dr. | de |
dc.date.examination | 2011-06-27 | de |
dc.subject.dnb | 570 Biowissenschaften | de |
dc.subject.dnb | Biologie | de |
dc.subject.gok | WF 200 | de |
dc.description.abstracteng | Selenocysteine is the 21st amino acid
which is incorporated into proteins by recoding a stop codon UGA
followed by a selenocysteine insertion sequence (SECIS) of the
mRNA. In bacteria, selenocysteine insertion requires specialized
machinery which includes selenocysteine-specific tRNASec,
translation factor SelB which delivers Sec-tRNASec to the ribosome,
as well as proteins SelA, SelD, and seryl-tRNA synthetase which are
required to produce Sec-tRNASec. The aim of this work is to develop
experimental assays to study Sec incorporation into proteins in
vivo and in vitro. As an in vivo assay, we designed the dual
luciferase reporter assay and validated its performance using
Western blots and luciferase reactions. The efficiency of Sec
incorporation was 35% independent of growth conditions. Rapidly
growing cells required additional selenium source for efficient Sec
insertion. This level of UGA recoding could be reproduced in the
fully reconstituted in vitro translation system upon synthesis of a
fragment of a natural selenoprotein FdfH. The recruitment of SelB
to the SECIS-element prior to translation prevented inhibition of
Sec insertion by RF2, a termination factor which usually recognizes
the UGA codon and competes with Sec-tRNASec for binding to the
ribosome. These results shed light on the importance of the SECIS
and on the mechanism by which a stop codon is redirected for
efficient readthrough by a specific tRNA. | de |
dc.contributor.coReferee | Stark, Holger Prof. Dr. | de |
dc.contributor.thirdReferee | Ficner, Ralf Prof. Dr. | de |
dc.subject.topic | Göttingen Graduate School for Neurosciences and Molecular Biosciences (GGNB) | de |
dc.subject.eng | selenocysteine | de |
dc.subject.eng | SECIS-element | de |
dc.subject.eng | in vivo | de |
dc.subject.eng | in vitro | de |
dc.subject.eng | translation | de |
dc.subject.eng | ribosome | de |
dc.subject.eng | selB | de |
dc.subject.bk | 42.00 | de |
dc.identifier.urn | urn:nbn:de:gbv:7-webdoc-3587-0 | de |
dc.identifier.purl | webdoc-3587 | de |
dc.affiliation.institute | Göttinger Graduiertenschule für Neurowissenschaften und Molekulare Biowissenschaften (GGNB) | de |
dc.identifier.ppn | 730380807 | de |