Characterizing the role of pmt-2 in C. elegans larval development
Disciplines
Biology | Genetics
Abstract (300 words maximum)
Nematode parasitic infections are becoming more prevalent in humans and animals. It is estimated that over 3 billion people have gotten at least one nematode infection in their lives, whether from hookworm, roundworm, or pinworm. Working on parasitic nematodes is difficult because they can be hard to culture in the lab. To circumvent this problem, we took the approach to working with Caenorhabditis elegans, a non-parasitic nematode that is easy to grow in culture. Studies have shown that nematodes synthesize phosphatidylcholine by a novel pathway which uses the enzymes PMT-1 and PMT-2. Our focus is to develop enzyme inhibitors of PMT-2, because the phosphatidylcholine biosynthetic pathway is crucial to the synthesis of membrane phospholipids. In particular, the PMT-2 enzyme is unique to nematodes, which should make it easier to target nematodes specifically. The pmt-2(ok2419) V/nT1[qIs51] strain is a homozygous lethal deletion mutation balanced by GFP-tagged translocation chromosome. Translocation balancers are difficult to work with because of aneuploidy, which is when there is an abnormal number of chromosomes in a cell. To facilitate our characterization of the pmt-2(ok2419) mutation, we set out to build a simple linked inversion balancer. Ten wildtype males were crossed with tmC16 homozygous mutants, then the resulting heterozygous F1 males crossed with oxTi396[eft-3p::mCherry] hermaphrodites. This cross was transferred daily to new plates to not have any self-fertilization with progeny in newer generations. These two reporters together are expected to have a mCherry marker in the pharynx and broad red nuclear fluorescence in body wall muscles and other tissues. Our preliminary data indicates that PMT-2 mutants may fail to molt out of the first larval stage. Work is ongoing to better characterize the role PMT-2 in larval development.
Academic department under which the project should be listed
CSM - Molecular and Cellular Biology
Primary Investigator (PI) Name
Martin Hudson
Characterizing the role of pmt-2 in C. elegans larval development
Nematode parasitic infections are becoming more prevalent in humans and animals. It is estimated that over 3 billion people have gotten at least one nematode infection in their lives, whether from hookworm, roundworm, or pinworm. Working on parasitic nematodes is difficult because they can be hard to culture in the lab. To circumvent this problem, we took the approach to working with Caenorhabditis elegans, a non-parasitic nematode that is easy to grow in culture. Studies have shown that nematodes synthesize phosphatidylcholine by a novel pathway which uses the enzymes PMT-1 and PMT-2. Our focus is to develop enzyme inhibitors of PMT-2, because the phosphatidylcholine biosynthetic pathway is crucial to the synthesis of membrane phospholipids. In particular, the PMT-2 enzyme is unique to nematodes, which should make it easier to target nematodes specifically. The pmt-2(ok2419) V/nT1[qIs51] strain is a homozygous lethal deletion mutation balanced by GFP-tagged translocation chromosome. Translocation balancers are difficult to work with because of aneuploidy, which is when there is an abnormal number of chromosomes in a cell. To facilitate our characterization of the pmt-2(ok2419) mutation, we set out to build a simple linked inversion balancer. Ten wildtype males were crossed with tmC16 homozygous mutants, then the resulting heterozygous F1 males crossed with oxTi396[eft-3p::mCherry] hermaphrodites. This cross was transferred daily to new plates to not have any self-fertilization with progeny in newer generations. These two reporters together are expected to have a mCherry marker in the pharynx and broad red nuclear fluorescence in body wall muscles and other tissues. Our preliminary data indicates that PMT-2 mutants may fail to molt out of the first larval stage. Work is ongoing to better characterize the role PMT-2 in larval development.