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Philipps-Universität Marburg
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Abstract
5‘ end maturation of precursor tRNAs is catalyzed by the enzyme Ribonuclease P (RNase P)
in all kingdoms of life. RNase P enzymes characterized in bacteria, archaea, yeast nuclei /
mitochondria and animal cell nuclei are ribozymes composed of one catalytic RNA subunit
and one to ten protein subunits. Recently, human mitochondrial RNase P was identified to be
composed of three polypeptides but to lack any RNA subunit. Thereupon, the RNase P in
land plants and kinetoplastids was found to consist of only one single polypeptide and was
hence named “PRORP” (proteinaceous RNase P).
In the hyperthermophilic bacterium Aquifex aeolicus, neither a gene for the RNA nor the
protein component of bacterial RNase P has been identified in its sequenced genome.
However, RNase P activity can be detected in A. aeolicus cell lysates. This PhD thesis aimed
at developing a robust and reproducible method for the purification and enrichment of A.
aeolicus RNase P activity. Mass spectrometry analysis identified one A. aeolicus protein in
highly purified fractions. According to homology structure modeling, this protein might well be
involved in A. aeolicus RNase P activity. Furthermore and for the first time, a protocol for
successful in vitro reconstitution of RNase P activity from RNA and protein preparations from
A. aeolicus fractions was established. The reconstitution experiments provide evidence that
RNA as well as protein components are involved in A. aeolicus RNase P activity. Protein and
RNA analysis also suggest an association of A. aeolicus RNase P with ribosomes.
Additionally, roughly 100 noncoding RNA (ncRNA) candidates, many represented by sense
and antisense transcripts, were identified bioinformatically and by deep sequencing (RNAseq);
selected ncRNA candidates were verified via Northern blot detection in total RNA of A.
aeolicus.
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Dates
Created: 2014Issued: 2015-06-15Updated: 2018-05-07
Faculty
Fachbereich Chemie
Publisher
Philipps-Universität Marburg
Language
ger
Data types
DoctoralThesis
Keywords
RNase PRNase PRibonucleoproteinesAquifex aeolicus
DFG-subjects
Endoribonuklease PRibonucleoproteineAquifex aeolicus
DDC-Numbers
570
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Nickel, Astrid Ingeborg: Aufreinigung und Charakterisierung der RNase P in Aquifex aeolicus. : Philipps-Universität Marburg 2015-06-15. DOI: https://doi.org/10.17192/z2014.0250.
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This item has been published with the following license: In Copyright