Transcription and translation of mammalian mitochondrial DNA (mtDNA) occurs within the mitochondrial matrix to produce oxidative phosphorylation subunits required for efficient energy production. These mtDNA-encoded subunits complex with mitochondrial-localized, nuclear-encoded subunits to form the respiratory chain, and aberrant production or function of these subunits can cause devastating human disease. In addition to 13 oxidative phosphorylation subunits, mtDNA encodes 2 rRNAs and 22 tRNAs. All proteins required for mitochondrial RNA transcription, processing, and translation are encoded in the nucleus and translocated into the mitochondria. Here, I characterize over 100 nuclear-encoded mitochondrial proteins with predicted RNA-binding domains. Using RNAi and an RNA profiling approach, MitoString, we further characterize previously identified RNA processing factors and identify the novel regulator FASTKD4, which influences the abundance of a subset of mitochondrial mRNAs. Next, we apply knowledge of the RNA degradation component SUPV3L1 gleaned from our RNAi studies and previous research to test whether a specific set of variants influence the function of this gene in patient fibroblasts. Using MitoString, we find no evidence of pathogenicity of these variants in our fibroblast model. Our approach highlights the value of a thorough understanding of mitochondrial proteins and the necessity of experimental techniques to validate the effect of variants found in exome-sequencing studies. Finally, we take an unbiased approach to characterizing the mitochondrial transcriptome of mouse liver by sequencing RNA from sequentially enriched mitochondrial fractions. Although we find an abundance of nuclear-encoded 5S rRNA, consistent with previous research, we fail to identify any imported nuclear-encoded tRNAs. Uniting genomics, biochemistry, and medicine, these findings advance our understanding of mitochondrial RNA biology.

Die vorliegende Arbeit soll dazu beitragen, Unstimmigkeiten in den Verwandtschaftsverhältnissen innerhalb der Papionini, einem Stamm innerhalb der Altweltaffen (Cercopithecidae), zu klären. Die Papionini, die zusammen mit den Cercopithecini die Unterfamilie der Cercopithecinae bilden, beinhalten sieben Gattungen (Macaca, Cercocebus, Mandrillus, Lophocebus, Papio, Theropithecus, Rungwecebus) und 45 Arten. Sechs der sieben Gattungen kommen heute hauptsächlich in Afrika vor. Eine Ausnahme ist die Gattung Papio, die mit einer Art (P. hamadryas) auch in Südwest-Arabien vorkommt. Im Gegensatz zu den sechs hauptsächlich afrikanischen Gattungen hat die siebte Gattung (Macaca) nur ein kleines Verbreitungsgebiet im Norden Afrikas und kommt sonst hauptsächlich in Asien vor. Fossilfunde belegen allerdings, dass während des Plio- und Pleistozäns die Gattungen Macaca und Theropithecus auch in Europa vorkamen. Von der Gattung Theropithecus, die heute ausschließlich in Afrika beheimatet ist, wurden zudem auch Fossilien aus dem Pliozän im Norden Indiens gefunden. Die Verwandtschaftsbeziehungen innerhalb der Papionini wurden bisher mit Hilfe morphologischer und genetischer Merkmale untersucht, allerdings waren die Ergebnisse nicht immer übereinstimmend und es gibt immer noch viele Unklarheiten. Zum einen ist nicht eindeutig geklärt, wie die Gattungen Papio, Lophocebus und Theropithecus zu einander in Beziehung stehen. Zum anderen ist auch unklar, wie die einzelnen Pavianarten innerhalb der Gattung Papio mit einander verwandt sind. Außerdem sind auch die Verwandtschaftsverhältnisse zwischen und innerhalb der Artgruppen der Makaken nicht eindeutig geklärt. Um mehr Klarheit in die Evolution der Papionini zu bringen, habe ich im Rahmen dieser Arbeit drei Studien durchgeführt (Kapitel 2-4). Ziel dabei war es, Verwandtschaftsbeziehungen auf unterschiedlichen taxonomischen Ebenen (zwischen und innerhalb von Gattungen, sowie innerhalb einer Art) zu untersuchen. Dazu wurden komplette mitochondriale Genome von Vertretern der Papionini sequenziert und damit Phylogenien und Aufspaltungszeiten berechnet. Die Ergebnisse meiner Arbeit zeigen unter anderem drei Hauptkladen innerhalb der Papionini (Kapitel 2): 1) Papio, Theropithecus, Lophocebus; 2) Mandrillus, Cercocebus; 3) Macaca, wobei Macaca in der mitochondrialen Phylogenie näher mit Mandrillus und Cercocebus verwandt zu seien scheint und nicht wie erwartet, als Schwestergruppe der afrikanischen Papionini abgebildet wird; ein Ergebnis, das im Widerspruch zu nukleären und morphologischen Studien steht. Meine Arbeit zeigt auch, dass komplette mitochondriale Genome in manchen Fällen nicht ausreichen, um phylogenetische Beziehungen vollständig zu rekonstruieren. So bleibt weiterhin unklar wie die Gattungen Papio, Theropithecus und Lophocebus zueinander stehen (Kapitel 2). Außerdem zeigen die Ergebnisse Paraphylien für Mandrillus und Cercocebus (Kapitel 2), sowie innerhalb der Paviane (Kapitel 3). Die Paviane werden dabei gemäß ihrer geographischen Verbreitung und nicht nach ihrer taxonomischen Zugehörigkeit abgebildet, wodurch die meisten Pavian-Arten paraphyletisch sind. Der Grund für diese Baumtopologie ist sehr wahrscheinlich sekundärer Genfluss zwischen parapatrisch vorkommenden Pavian-Arten. In der dritten Studie (Kapitel 4), in der innerartliche Verwandtschaftsverhältnisse innerhalb einer südostasiatischen Makaken-Artgruppe (Macaca fascicularis) untersucht wurden, zeigt sich eine klare Unterteilung in eine kontinentale und eine insulare Klade. Sowohl die kontinentale, als auch die insulare Linie sind auf Sumatra zu finden, was für einen sekundären genetischen Austausch zwischen beiden Populationen spricht. Generell kann man sagen, dass komplette mitochondriale Genome robuste Phylogenien mit hoher statistischer Unterstützung ergeben, die eine gute Grundlage für künftige vergleichende Studien bilden. Die berechneten Aufspaltungszeiten stimmen weitestgehend mit vorherigen Studien überein, wobei sich die ermittelten Konfidenzintervalle verkleinert haben. Allerdings zeigt die Arbeit auch, dass Phylogenien basierend auf mitochondrialen Genomen keine hohe Auflösung erzielen wenn sich Taxa innerhalb kurzer Zeit voneinander trennten. Die hier gezeigten Paraphylien und die abweichenden Ergebnisse zu nukleären Studien wurden höchstwahrscheinlich durch sekundären genetischen Austausch hervorgerufen. Um Verwandtschaftsverhältnisse möglichst exakt rekonstruieren zu können, müssen neben der maternal-vererbten, mitochondrialen Linie noch paternal- und biparentalvererbte Merkmale in Betracht gezogen werden. Zu beachten ist in diesem Zusammenhang, dass ein bestimmter molekularer Marker immer nur eine mögliche Phylogenie von vielen wiedergibt.

The human mitochondrial DNA polymerase (Pol γ) catalyzes mitochondrial DNA synthesis, and thus is essential for the integrity of the organelle. Mutations of Pol γ have been implicated in more than 150 human diseases. Reduced Pol γ activity caused by inhibition of anti-HIV drugs targeted to HIV reverse transcriptase confers major drug toxicity. To illustrate the structural basis for mtDNA replication and facilitate rational design of antiviral drugs, I have determined the crystal structure of human Pol γ holoenzyme. The structure reveals heterotrimer architecture of Pol γ holoenzyme with a monomeric catalytic subunit Pol γA, and a dimeric processivity factor Pol γB. While the polymerase and exonuclease domains in Pol γA present high structural homology with the other members of the DNA Pol I family, the spacer between the two functional domains shows a unique fold, and constitutes the subunit interface. The structure suggests a novel mechanism for Pol γ’s high processivity of DNA replication. Furthermore, the structure reveals dissimilarity in the active sites between Pol γ and HIV RT, thereby indicating an exploitable space for design of less toxic anti-HIV drugs. Interestingly, the structure shows an asymmetric subunit interaction, that is, one monomer of dimeric Pol γB primarily participates in interactions with Pol γA. To understand the roles of each Pol γB monomer, I generated a monomeric human Pol γB variant by disrupting the dimeric interface of the subunit. Comparative studies of this variant and dimeric wild-type Pol γB reveal that each monomer in the dimeric Pol γB makes a distinct contribution to processivity: one monomer (proximal to Pol γA) increases DNA binding affinity whereas the other monomer (distal to Pol γA) enhances the rate of polymerization. The pol γ holoenzyme structure also gives a rationale to establish the genotypic-phenotypic relationship of many disease-implicated mutations, especially for those located outside of the conserved pol or exo domains. Using the structure as a guide, I characterized a substitution of Pol γA residue R232 that is located at the subunit interface but far from either active sites. Kinetic analyses reveal that the mutation has no effect on intrinsic Pol γA activity, but shows functional defects in the holoenzyme, including decreased polymerase activity and increased exonuclease activity, as well as reduced discrimination between mismatched and corrected base pair. Results provide a molecular rationale for the Pol γA-R232 substitution mediated mitochondrial diseases. / text

Pollination, one of our ecosystem services, is considered to be in critical condition due to a worldwide reduction in pollinators and their biodiversity. As the agricultural landscape becomes more and more intense, the pollinators lose important food and living resources.     In temperate ecosystems, bumblebees (Bombus spp) are an important group of wild pollinators, and as with pollinators in general, they are declining in both abundance and richness, in Sweden as well as other countries.      The purpose of this study was to see if bumblebee species number of a location is linked to the location’s number of flowering angiosperm species in northwestern Scania when examining eight locations, and to see if gene flow existed between four chosen locations.        The result of this study suggests that it is not possible to tell from the flowering angiosperm species how many bumblebee species that will be abundant, but that it might be possible to tell the number of bumblebee individuals. With the number of bumblebee species, the abundant Fabaceae species was more important than the total number of flowering angiosperms of the location. The number of abundant Fabaceae species was strongly correlated to the bumblebee diversity index of the locations, indicating that it is a group of flowers closely linked to bumblebees.      To see if gene flow occurred between the chosen locations, mtDNA sequences were compared in neighbor joining trees. The result showed that though some tendencies of isolation existed, gene flow seemed to occur in general between the locations in that fragmented and human dominated landscape of northwestern Scania.

An almost unbroken fringing reef runs along the east coast of Africa, the lagoon inside the reef is the foundation of almost all artisanal fisheries. It is a low-tech fishery conducted by many people. Some areas can have up to 19 fishermen per square kilometer. High fishing pressures, coupled with declining fish stocks has led to changes in mean size and reproductive age of many exploited species. There is a vital and urgent need for scientifically based management systems, including the utilization of genetic information to guide management practices. This thesis aims to investigate the presence of genetic structures in the western Indian Ocean. In order to do that we first investigated the historical patterns of connectivity throughout the region (paper I). In papers II and III we focused on local scale connectivity in Kenya and Tanzania and finally in paper IV we investigate the large-scale contemporary gene flow throughout the Western Indian Ocean. In paper III we also investigate the temporal genetic variation at one site and compare it to the small-scale genetic variation along a stretch of the Kenyan coastline. Some overall conclusions that can be drawn from my body of work are: there are genetic structures present in the western Indian Ocean even though the apparent lack of physical barriers. Major oceanic currents aid evolutionary dispersal patterns. A single geographic site need not be genetically homogenous or temporally stable. Island sites are genetically more homogenous than mainland sites. In conclusion, there are clear and distinct genetic structures present especially in Siganus sutor, the most targeted fish for the artisanal fishery in East Africa. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.</p>

ABSTRACT MORPHOMETRIC, MtDNA AND MICROSATELLITE ANALYSIS IN HONEYBEE POPULATIONS (Apis mellifera L.) OF NORTH AND NORTHWEST IRAN Morphometric measurements, mitochondrial DNA analyses and 5 microsatellite loci were used to investigate variation in the honeybee populations of Iran and comparing it with the Turkish populations. Five honeybee populations were sampled from North and west north of Iran. In morphometric aspect of the study 23 characters were measured from left forewings and hindlegs of honey bee samples. The data were analysed by multivariate statistical analyses. By using mtDNA analyses length polymorphism of the intergenic region COI-COII of mitochondrial DNA was studied. After amplification of this region by the polymerase chain reaction, DraI enzyme was used for restriction of amplified region. Results of mtDNA studies show no diversity between four populations and all of them exhibit the same C1 pattern. Five microsatellite loci (A7, A24, A28, A43 and A113) were used in this studies.A high level of average heterozygosity changing between 0.611 and 0.709 was detected in Iranian honey bee populations, and a significant degree of polymorphism was observed. Although Urmia, Sarein and Viladereg populations are similar, Amol population which has located in northern Iran shows a significant difference from other populations. Result obtained form morphometric studies are supporting microsatellite analyses. By comparing data obtained form Iranian honey bee populations with Turkish population (Hakkari), western populations (Urmia, Sarein and Viladereg) are more similar to Hakkari population. It is found Amol is significantly different form other populations and better represents Iranian honeybee.

Kangal dogs are the most popular dogs of Turkey due to their strength, intelligence, loyalty, endurance to extreme temperatures and their lack of predatory behavior towards livestock. In this study to provide genetic information about the distinctness of Kangal and Akbash dogs and hence to provide a basis to conserve them separately, 585 base pair of the mitochondrial DNA (mtDNA) control region sequence was analysed in 105 Kangal and 9 Akbash dog samples. All the results indicated that Kangal and Akbash dogs were different from each other. Comparison of the Turkish data with those from other dogs revealed that Kangal dogs harbour a rare haplogroup which is only seen in Scandinavian (36%), Portuguese (20%), Turkish (20%) dogs and only one Spanish dog, but not in Akbash, Middle Eastern, other European, Eastern Asian and Indian dogs. Furthermore, comparison of the Kangal and Akbash dogs with the dogs from different geographic regions indicated that Kangal dogs are genetically closer to Scandinavian and South West Asian dogs whereas Akbash dogs are more similar to European and East Asian dogs, based on the mtDNA control region sequences Today, the sizes of Kangal and especially Akbash populations are decreasing. An urgent program for their conservation is needed. In order to conserve them separately, it must be understood that these two dogs are genetically distinct. That is why, the main purpose of the present study is to provide genetic information about the distinctness of Kangal and Akbash dogs.

Coral reefs constitute some of the most prolific and diverse ecosystems on our planet, but also among the most threatened. This thesis investigates the effects of environmental stressors on corals’ metabolism and symbiont diversity. Paper I shows that the coral Turbinaria mesenterina withstood a single stressor while a combination of two stressors (decreased salinity and increased seawater temperature) lead to decreased metabolism. Increased seawater temperature in combination with two stressors (enhanced nutrients and decreased salinity) lead to rapid mortality of all specimens. Paper II shows that chronic stress in combination with increased seawater temperature affects coral species differently. Porites lutea did not show any difference in response to temperature increase, regardless of environmental disturbance history, while Galaxea fascicularis’ metabolism was negatively affected in chronically disturbed corals but not in corals from less disturbed areas. The main explanation for the difference in response between the two species is different compositions of endosymbionts as found in paper III. P. lutea only harboured the symbiont C15, regardless of environment, whilst D1a dominated the nearshore G. fascicularis and C1 dominated offshore corals. In paper IV there was a clear inshore-offshore pattern of D1a along the whole coast of Vietnam, where D1a dominated inshore. In contrast, the five symbionts belonging to group C displayed a strong latitudinal gradient, with diversity increasing from north to south. The coral host showed higher diversity offshore than inshore. The thesis emphasizes the importance of improving water quality (paper I and II) and protecting marginal areas since tolerant coral hosts and symbionts can be found there (paper III and IV), as well as safeguarding areas with high symbiont diversity (paper IV) to increase the ability of corals to withstand future environmental changes. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.

The European rabbit, Oryctolagus cuniculus, was introduced to Australia in 1859 and quickly became a significant vertebrate pest species in the country across a wide distribution. In arid and semi-arid environments, rabbit populations exist as metapopulations - undergoing frequent extinction recolonisation cycles. Previous studies identified population genetic structuring at the regional level between arid and semi-arid environments, and habitat heterogeneity was suggested as a possible causal factor. For the most part, rabbit behaviour has been overlooked as a factor that could contribute to explaining population genetic structure in arid and semi-arid environments. This study utilised a combination of genetic sampling techniques and a simulated territorial intrusion approach to observing wild rabbit behaviour in arid and semi-arid environments. The genetic component of the study compared population samples from each region using four polymorphic microsatellite loci. The behavioural component examined variation in the level of territoriality exhibited by three study populations in the arid region towards rabbits of known versus unknown origins (resident vs transgressor (simulating dispersal)). A difference was observed in population genetic structure determined from nuclear markers between arid and semi-arid regions, which supports findings of previous research using mitochondrial DNA data in the same area. Additionally, differences in aggressive response to known vs unknown rabbits were identified in parts of the arid region, which together with the effects of habitat heterogeneity and connectivity may explain the observed differences in population genetic structure. Knowledge of behavioural plasticity and its effect on relative dispersal success and population genetic structure may contribute to improved management and control of feral rabbit populations at the regional level within Australia; and may assist with conservation efforts in the species' natural range in Europe.

The influence of genotype variation on phenotype has been a longstanding question in biology but it is now one of the greatest challenges of the post-genomics era. Discovering the link between common gene variants that affect phenotypes within and between populations is likely to provide insight into the molecular physiology of phenotypic traits and the mechanisms by which they evolve. The overall goal of this thesis is to link naturally occurring genotypic variation with the organism??s phenotype. The specific goal of this thesis is to connect natural variation in the mitochondrial genotype with the organismal phenotype using the model organism Drosophila simulans. Mitochondria are intracellular organelles found in most eukaryotes and produce over 90% of the energy needed by cells. Determining the connection of mitochondrial genotype to whole organism phenotype is of particular interest because of the broad use of mitochondrial (mt) DNA as a molecular marker in evolutionary biology and population genetics, the organelle??s central role in cellular energy production, the potential for the mitochondria to influence organismal distribution particularly in the face of climate change and in human degenerative disease. I use the model organism D. simulans because it has high genetic variability, can be easily sampled from the wild and manipulated in the lab, and the energy producing reactions that take place in its mitochondria are highly conserved among metazoa. I studied naturally occurring mutations to understand the influence of these changes in natural populations. The four studies in this thesis have employed a Genotype-Biochemistry-Phenotype (GBP) model to link naturally occurring variation in the mitochondrial genotype with organism phenotype in D. simulans mitochondria. Three major conclusions can be drawn from the thesis that follow the genotype to biochemistry to phenotype model. Firstly, a subset of the mutations in genes that comprise the mitochondrial genotype is functionally significant. Secondly, the biochemical efficiency of OXPHOS is regulated by mitochondrial homeostasis. Thirdly, key organismal life history traits influenced by the mitochondrial genotype and this is mediated through the biochemistry of OXPHOS.