Graužikų naujų poliomos virusų koduojamų baltymų sintezės bei jų funkcijos tyrimas bakterijų ir mielių ląstelėse /

VILNIUS UNIVERSITY LIFE SCIENCES CENTRE Justė Remeikaitė Bachelor thesis Investigating the Synthesis and Function of Proteins Encoded by Novel Rodent Polyomaviruses in Bacterial and Yeast Cells SUMMARY Polyomaviruses (PyVs) belong to the Polyomaviridae family of small, non–enveloped viruses, whose genome is composed of circular dsDNA packaged into icosahedral particles consisting of 3 structural proteins: VP1, VP2, VP3. These viruses infect mammals, avian, fish and in most cases are host specific. The ICTV (International Committee on Taxonomy of Viruses) has currently confirmed 117 species of polyomaviruses, 16 of which infect rodents (Rodentia). The discovery of novel PyV species and the characterization of their genome may help to understand the evolution of these viruses and their relationship to the host. In this study, the proteins of MarPyV2 and CriPyV, novel rodent polyoma viruses, and their synthesis in bacterial and yeast cells were studied. The genomes of MarPyV2 and CriPyV contain an additional open reading frame encoded in a non–coding regulatory region. These proteins are successfully synthesised in bacterial and yeast cells and are also used for MAb production. The resulting MAb are antigen–specific and suitable for the detection of these proteins. The regulatory proteins encoded by MarPyV2, STAg and LTAg, were synthesised in E. Coli bacteria and Saccharomyces cerevisiae yeast cells. Successful protein synthesis in bacteria was observed for the STAg protein and LTAg DnaJ domain. RNA and protein synthesis of the regulatory genes encoded by MarPyV2 in yeast was tested in two steps: protein translation and RNA transcription analyses. In yeast and bacterial cells, protein synthesis analyses have revealed the expression of STAg and LTAg proteins, but full–length LTAg or LTAg without the intron (cLTAg) proteins are not efficiently synthesised. Application of molecular tools reveals that the intronless LTAg, LTAg and STAg genes are successfully transcribed in yeast cells and inefficient transcript splicing of the LTAg gene occurs. Further characterisation of unknown function viral proteins requires the identification of protein domains and whether they interact with DNA or the host immune system. The identification of non–silent amino acid mutations would suggest strong positive selection and could indicate rapid evolution of the genome along with the host.

Grantor dc:publisher

Institutional Repository of Vilnius University

Year dc:date

2024

Author dc:creator

• Remeikaitė, Justė,

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

lit

Repository record dc:identifier

https://repository.vu.lt/VU:ELABAETD210640018&prefLang=en_US

OAI identifier oai:identifier

oai:vu.lt:elaba:210640018