Interaction analysis between proteins encoded by genes involved in growth and differentiation in Arabidopsis thaliana

Postgraduate Thesis uoadl:1318503 157 Read counter

ΠΜΣ Μικροβιακή Βιοτεχνολογία
Library of the School of Science
Deposit date:
Πίκουλα Λαμπρινή
Supervisors info:
Επικ. Καθηγητής Κ. Χαραλαμπίδης (επιβλέπων), Επικ. Καθηγητής Α. Ρούσσης, Επικ. Καθηγητής Δ. Χατζηνικολάου
Original Title:
Μελέτη πρωτεϊνικών αλληλεπιδράσεων γονιδίων που εμπλέκονται στην ανάπτυξη και διαφοροποίηση του Arabidopsis thaliana
Translated title:
Interaction analysis between proteins encoded by genes involved in growth and differentiation in Arabidopsis thaliana
The objective of this study was the analysis of predicted interactions between
proteins involved in developmental processes in Arabidopsis thaliana, including
embryogenesis and formation of shoot and root apical meristems which induce the
generation of all the aerial and subterranean plant organs.
Protein-protein interactions are intrinsic to virtually every cellular process
ranging from DNA replication to metabolism, formation of cellular
macrostructures and enzymatic complexes. Apart from the evident structural
requirements provided by a plethora of protein-protein interactions, there is a
large number of transient protein-protein interactions, which are equally
important for the regulation of cellular processes. Post-translational
modifications of proteins involve such transient interactions between the
enzymes (e.g. kinases, phosphatases, acyl transferases) and their protein
substrates. To date, various methods have been developed for the investigation
of protein-protein interactions, while each of the approaches has its own
strengths and weaknesses, especially with regard to the sensitivity and
specificity of the method. The yeast-two-hybrid system (Y2H) is a widely spread
technique used for the identification of new protein interactions, the
verification of predicted interactions and the determination of protein
domains. The Y2H assay was performed in this study in order to reveal useful
data for the functional characterization of genes At5g14540 (UBA-like),
At5g14520 (AtPES), At5g14510 (Arm), At5g14530 (WD1) and At5g66240 (WD2). These
genes were isolated and examined for interactions of their products with
proteins encoded by the following genes of known function: VIP3, DDB1a, DDB1b,
ASK1 and ELP3. The selection of all genes in the assay was based on existing
studies according to which the domains included in their sequences participate
in crucial interactions for the construction of dynamic protein complexes and
also on functional predictions from the genes databases. The analysis of 33
protein-protein interactions revealed 5 positive results. According to these
results the UBA-like protein interacts with ΑRM, WD2, DDB1A and ASK1 proteins.
This study also confirmed the known interaction between proteins VIP3 and
DDB1A, which was used as a positive control in the interaction analysis. The
interactions of UBA-like protein with DDB1A and ASK1 may involve UBA-like in
the process of protein degradation through the ubiquitin 26S proteasome
pathway. However, these two interactions may be parts of the construction of
non-proteolytic complexes. Furthermore, the interaction of UBA-like with ARM
protein probably indicates an indirect connection of ARM with DDB1A and ASK1 in
ubiquitin ligase complexes, while the results of interaction analysis between
these proteins showed up negative. Finally, the interaction of UBA-like with
WD2 protein implies the participation of WD2 as a subunit in a CUL4-DDB1-DCAF
ubiquitin ligase complex and it also explains the pleiotropic phenotype of wd2
mutants. Further experimental approaches, such as co-immunoprecipitation
experiments are necessary for the verification of all the above interactions,
as well as for the complete understanding of their functional roles.
Arabidopsis thaliana, Protein protein interactions, Yeast two hybrid, Genes, Ubiquitin ligase complex
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