Pre-mRNA splicing: regulation of monoamine oxidase B and oxygen-dependent genes expression

Author, Institution: Eglė Jakubauskienė, Kaunas University of Technology

Science Area, Field of Science: Technological Sciences, Chemical Engineering – 05T

Scientific Supervisor:  dr. Arvydas KANOPKA (Vilnius University, Biomedical Sciences, Biology, 01B).

Dissertation Defence Board of Chemical Engineering Science Field:

Prof. Dr. Petras Rimantas VENSKUTONIS (Kaunas University of Technology, Technological Sciences, Chemical Engineering, 05T) – chairman;
Prof. Dr. Darius BALČIŪNAS (Temple University, USA, Biomedical Sciences, Medicine, 07B);
Dr. Ilona JONUŠKIENĖ (Kaunas University of Technology, Technological Sciences, Chemical Engineering, 05T);
Prof. Dr. Kęstutis SUŢIEDĖLIS (National Cancer Institute, Biomedical Sciences, Biology, 01B);
Prof. Dr. Dalė VIEŢELIENĖ (Lithuanian University of Health Sciences, Biomedical Sciences, Biology, 01B).

The Doctoral Thesis is available at the library of Kaunas University of Technology (K. Donelaičio St. 20, Kaunas)

Annotation:

Pre-mRNA splicing is one of the essential steps in the regulation of the eukaryotic gene expression, and it is not surprising that the disruption of the normal splicing patterns can lead to various human diseases or contribute to their severity.

Aim

1. To analyze the effect of G/A polymorphism in MAO-B gene intron 13 sequence for pre-mRNA splicing, to identify the factors involved in such regulation and to study the MAO-B protein expression levels in the platelet fraction of healthy individuals and Parkinsonian patients.
2. To identify factors regulating oxygen tension-dependent alternative pre-mRNA splicing and to find a possibility to control the hypoxia-dependent splice site usage.

This study demonstrates that G/A polymorphism in MAO-B gene intron 13 sequence stimulates the efficiency of the intronic sequence removal from MAO-B pre-mRNA and that it influences protein expression levels in cells. The increased MAO-B protein levels could serve as a Parkinson’s disease marker.

Alternative pre-mRNA splicing plays an important role in cellular response to hypoxia producing multiple mRNA isoforms from the same pre-mRNA. We report that SR proteins in hypoxic cells are hyper-phosphorylated in comparison with normoxic cells, and this enhancement is caused by an increase in the expression of specific SR protein kinases (SRPK1, SRPK2 and CLK1) in hypoxic cells. The reduction of CLK1 cellular expression levels reduces the hypoxia-dependent CAIX and Cyr61 alternative pre-mRNA splicing.