Genetsko ozadje uravnavanja biogeneze mitohondrijev

Renčelj, Andrej (author), Dovč, Peter (mentor)

Biogeneza mitohondrijev je zapleten proces, v katerem je potrebno usklajeno izražanje več kot 1500 genov. V doktorski nalogi smo pripravili profil izražanja nekaterih pomembnih genov, ki so vpleteni v regulacijo biogeneze mitohondrijev. NRF-1, NRF-2, PPARGC1A, TFAM, POLG, POLRMT in TWNIKLE so kandidatni geni, ki s svojim nepravilnim delovanjem lahko močno vplivajo na biogenezo in delovanje mitohondrijev. Profil izražanja izbranih genov smo preverili v različnih tkivih (m. semispinalis capitis, m. semimembranosus, srčna mišica, možgani, vranica in jetra) pri prašičih moškega spola hibrida 1244 in avtohtone slovenske pasme krško poljski prašič. Obenem smo preverili še profil izražanja krajše, alternativno izrezane oblike gena TFAM, ki je ključni transkripcijski faktor v mitohondriju. Iz podatkovnih zbirk ENSEMBL in NCBI smo pridobili do sedaj znane informacije o kandidatnih genih, ki so vpleteni v biogenezo mitohondrijev pri prašiču. Da smo lahko pripravili profil izražanja kandidatnih genov, smo uporabili metodo qPCR. Analiza podatkov je pokazala, da se krajša oblika gena TFAM (proteinu TFAM manjka HMG enota 1, ki je kodirana v eksonu 4) izraža v vseh izbranih tkivih. Domnevamo, da ima tak protein slabšo vezavno afiniteto. Z bioinformacijskimi metodami in statističnimi izračuni smo dokazali, da je PPARGC1A eden glavnih akterjev v začetnih fazah mitohondrijske biogeneze. Z analizo regulatornih poti smo lahko dokazali, da so PPARGC1A, NRF1 in SIRT1 močno medsebojno povezani pri uravnavanju biogeneze mitohondrijev, medtem ko neposredne povezave teh faktorjev s POLG, LONP1, OMA1 in TWNIKLE nismo mogli dokazati.

• Language: Slovenian
• Keywords: prašiči, mitohondriji, biogeneza mitohondrijev, alternativno izrezovanje, izražanje genov
• Work type: Doctoral dissertation
• Typology: 2.08 - Doctoral Dissertation
• Organization: BF - Biotechnical Faculty
• Year: 2017
• PID: 20.500.12556/RUL-98455
• COBISS.SI-ID: 4031624
• Publication date in RUL: 02.12.2017

Secondary language

• Language: English
• Title: Genetic architecture of mitochondrial biogenesis

Abstract

Mitochondrial biogenesis is complex process where expression of more than 1500 genes is necessary. The objective of this dissertation was developing expression profile of some important genes involved in regulation of mitochondrial biogenesis. NRF-1, NRF-2, PPARGC1A, TFAM, POLG, POLRMT, TWNIKLE are genes that influence on biogenesis and function of mitochondrion. For this research we chose six pig tissue (m. semispinalis capitis, m. semimembranosus, heart muscle, brain, spleen and liver). All pigs were male hybrid 1244 and krško polje breed. At the same time we have developed expression profile of alternatively spliced form of TFAM gene, which is important transcription factor in mitochondrial biogenesis. We used ENSEMBL and NCBI databases for collection of known pig data. For expression profile we used absolute quantification qPCR method. Analysis have shown that shorter, alternatively spliced, form of TFAM is expressed in all selected tissues. Shorter form of TFAM is missing HMG box 1 which is coded in exon 4 of the gene. From data obtained we conclude that this protein has impaired binding to mtDNA. With bioinformatics and statistical analysis we have shown that PPARGC1A is one of the most important proteins in first phases of mitochondrial biogenesis. With regulatory pathway analysis we have shown that PPARGC1A, NRF1 and SIRT1 are strongly connected in this process. For POLG, LONP1, OMA1 and TWNIKLE we could not prove that they were directly related to PPARGC1A, NRF11 and SIRT1.

• Keywords: pigs, mitochondrion, mitochondrial biogenesis, alternative splicing, gene expression