The long-term goal of this research is to determine how chromosome synapsis functions to promote proper chromosome segregation during meiosis. Chromosome missegregation during meiosis is directly tied to human infertility and is also the leading known genetic cause for mental retardation and developmental disabilities. Elucidating the basic mechanisms underlying proper chromosome segregation during meiosis will enable greater understanding of the intricate pathways that contribute to normal gametogenesis and fertility. During prophase I, homologous chromosomes pair and then synapse. Synapsis occurs via the assembly of a proteinaceous structure known as the synaptonemal complex that forms between homologous chromosomes. Successful assembly of the synaptonemal complex is a key prerequisite to proper chromosome segregation during meiosis. However, many basic questions about the kinetics of assembly of these structures remain unanswered. Our objective for this proposal is to determine how the process of synaptonemal complex assembly contributes towards its dual function of 1) maintaining a tight association between homologs and 2) promoting crossing over and its regulation.
Our first aim uses fast, live, 3-D fluorescence imaging and quantitative image analysis to determine the kinetics of synaptonemal complex assembly in budding yeast to answer several important questions. What is the rate of synapsis polymerization? Is it bidirectional or unidirectional? How far can synapsis extend from one initiation site? In the past, the answers of these questions have eluded investigation, due to the fact that in most organisms, multiple moving chromosomes are synapsing from a large number of sites, over a long time frame, in a highly compacted nucleus. To reduce the complexity of the problem, we propose to introduce a zip3 mutation that 1) limits the number of synapsing chromosomes to as low as one and 2) changes nucleation from multiple sites to one, or at most two sites, along the chromosome. Synapsis will be followed by imaging the Zip1 protein that has been previously coupled to GFP and used successfully to image the motion of fully synapsed chromosomes but not synapsis formation.
Our second aim will be to characterize the process of nucleation. To accomplish this task, we will couple components of the initiation complex to a ligand binding domain of the estrogen receptor that keeps the fused protein inactive until introduction of estrogen. We then can investigate how the introduction and timing of various known components of the initiation complex influences the progression of synapsis. For our last aim, we will determine whether changes in synapsis nucleation and polymerization rates affect crossing over and its regulation. Using a genome-wide approach developed in my lab for looking at crossover control in a single cell that has undergone meiosis, we will assess how particular changes in synaptonemal complex assembly and nucleation can affect crossover distribution and thus chromosome segregation.)

Public Health Relevance

Chromosome missegregation during meiosis is directly tied to human infertility and is also the leading known genetic cause for mental retardation and developmental disabilities. This work investigates the mechanisms in place to ensure faithful chromosome segregation by elucidating how the assembly of the synaptonemal complex contributes to this process. Such research may lead to new ideas for treatment of infertility or to development of diagnostic tests to detect potential problems of chromosome segregation early on before expensive medical and surgical treatments are attempted.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM097213-04
Application #
8616074
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Carter, Anthony D
Project Start
2011-03-15
Project End
2016-02-29
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
4
Fiscal Year
2014
Total Cost
$292,778
Indirect Cost
$103,278
Name
University of California San Francisco
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
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