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Identifier 000408567
Title In vitro μελέτη της πρωτεΐνης Enhancer of RNA interference 1 της Arabidopsis thaliana
Alternative Title In vitro study of Enhancer of RNA interference 1 homologue of Arabidopsis thaliana
Author Βλατάκης, Ιωάννης Χ
Thesis advisor Καλαντίδης, Κρίτων
Researcher Κοτσαμπάσης, Κυριάκος
Τσαγρή, Ευθυμία
Χαλεπάκης, Γεώργιος
Μπουριώτης, Βασίλης
Κοκκινίδης, Μιχαήλ
Πετράτος, Κυριάκος
Abstract ERI1 exonucleases are an important subgroup of 3’ exonucleases that participate and regulate the maturation of 3’ end of coding and non coding RNAs. In eykaryotes it has been proposed an evolutionary relationship between ERI1 functions and siRNA pathways. Study of plant homologues from Arabidopsis thaliana and Nicotiana benthamiana revealed their involvement in maturation of the 3’ end of chloroplastic rRNAs. In order to obtain additional information of the activitiy of plant homologues, a biochemical analysis of the A.thaliana homologue, AtERIL1, was performed. AtERIL1 retains 3’ exonuclease activity with a preference towards small RNA oligonucleotides. With less efficiency it can hydrolyse small DNA oligonucleotides and RNA oligonucleotides having cytosine to the 3’ end. Despite the lack of a conserved nucleic acid binding domain, AtERIL1 has the ability to bind to 5S rRNA precusors, small single stranded and double strounded RNA and DNA oligonucleotides. The binding capacity of AtERIL1 is affected by two lysine residues present before the EXOIII domain in the primary protein sequence. Direct activity towards rRNA precursors was not observed during in vitro experiments. This may be explained by either an indirect effect of ΑtERIL1 to rRNA chloroplastic maturation or by the possibility that AtERIL1 may process rRNA precursors as part of complexes consisting by protein or RNA factors. Furthermore, it was found that AtERIL1 was not able to functionally complement the bacterial homologue RNAse T in vivo. Lastly, based on co-immunoprecipitation experiments certain tRNA species were screened by northern blots for their accumulation and maturation status in N.benthamiana lines that either overexpress or downregulate ERI1 homologue. The results of the current thesis confirm the conservation of biochemical properties of ERI1 subgroup in plants and pave the way for hypotheses around its in vivo activities concerning both the mechanism and the type of substrates that may be processed by AtERIL1 in chloroplasts.
Language Greek
Subject 3' exonuclease
3' εξωνουκλεάση
Chloroplast
RNA exonuclease
RNA εξωνουκλεάση
Small RNA
Μικρά RNA
Χλωροπλάστες
Issue date 2017-05-25
Collection   Faculty/Department--Faculty of Sciences and Engineering--Department of Biology--Doctoral theses
  Type of Work--Doctoral theses
Permanent Link https://elocus.lib.uoc.gr//dlib/3/f/b/metadata-dlib-1493108368-578086-21481.tkl Bookmark and Share
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