Broad long-term objectives:
In Specific Aim 1, we will determine the phenotypes (e.g. 7 mitochondrially targeted drugs; non-mitochondrially targeted drugs; growth at low and high temperature) of 98 L-A strain pairs. We will identify strain pairs with strong L-A-dependent phenotypes; that is, N1/N1 ?1 L-A0 ? N1/N1 ?1 L-A+. We will also identify candidate QTGs for analysis in Aim 3.
In Specific Aim 2, we will determine the phenotypes (e.g. 7 mitochondrially targeted drugs; non-mitochondrially targeted drugs; growth at low and high temperature) of iso- nuclear pairs of F1; for example, N1 ?1 ? N2 ?0 ? N1/N2 ?1 and N1 ?0 ? N2 ?2 ? N1/N2 ?2. We will identify iso-nuclear F1 diploid strain pairs with strong mitochondrial genome-dependent phenotypes; that is, N1/N2 ?1 ? N1/N2 ?2. We will cross strains with strong mitochondrial genome-dependent phenotypes; for example, N1 ?1 ? N2 ?2 ? F1 N1/N2 (? recombinants). In multiple F1 N1/N2 (? recombinants) diploids, we will identify phenotypically relevant mitochondrial (?) QTG(s).
In Specific Aim 3 A, using our results from Aim 1 and 2, we will perform RNA-Seq on ethanol and dextrose grown isogenic L-A parent strains and L-A- and ? genotype-controlled iso-nuclear F1. We will identify L-A and ? genotype-dependent effects on the transcriptomes and potential candidate QTGs, which will aid downstream analysis.
In Specific Aim 3 B, we will (separately) sporulate multiple iso-nuclear F1 strain pairs with strong L-A- and/or mitochondrial genome-dependent phenotypes (identified in Aim 1, 2; RNA-Seq in Aim 3B) to generate L-A- and ? genotype-controlled F12 populations. In multiple L-A F12 ?1 vs. L-A F12 ?2 population pairs, we will identify phenotypically relevant nuclear QTGs. We will primarily focus on L-A and ? genotype-specific nuclear QTGs to advance our understanding of missing heritability, host-virus, and N-? interactions.

Public Health Relevance

In all species of eukaryotes, the mitochondrial and nuclear genomes are inherited in different manners. In all species of eukaryotes, the mitochondrial genome, which is physically small with relatively few genes, is required for respiration. In all species of eukaryotes, the bulk of the mitochondrial proteome is encoded by the nuclear genome and all mitochondrially-encoded genes must form complexes with nuclearly encoded genes for (proper) function. Despite the importance of the mitochondrial genome, the impact of mitochondrial polymorphisms on complex (i.e. quantitative/multi-genic) traits and phenotypes is ignored or poorly understood. Improving our understanding of mitochondrial genome polymorphisms and their impact on complex traits and phenotypes is aided by tractable model systems, such as S. cerevisiae. Much the same can be said of S. cerevisiae viruses and host-virus interactions. The health relatedness of this project lies in our study in S. cerevisiae of host-virus and nucleo- mitochondrial complex traits, which have been poorly understood, where they have been studied at all. Our study in S. cerevisiae of host-virus and nucleo-mitochondrial complex traits will aid our understanding of complex traits in general and of host-virus and nucleo- mitochondrial genetics in particular.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM118936-04
Application #
9939617
Study Section
Genetic Variation and Evolution Study Section (GVE)
Program Officer
Krasnewich, Donna M
Project Start
2017-08-16
Project End
2021-05-31
Budget Start
2020-06-01
Budget End
2021-05-31
Support Year
4
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Duke University
Department
Genetics
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705