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Biology subjects

Sean M Burgess

Publications and source records attributed to Sean M Burgess.

3 recordsLinked to original sources

An autonomous meiosis-specific region of yeast Nup2 (hNup50) promotes normal meiotic chromosome dynamics in Saccharomyces cerevisiae.

Meiosis is a specialized cellular program required to create haploid gametes from diploid parent cells. Prior to the first meiotic division, homologous chromosomes pair, synapse, and recombine to ensure their proper disjunction at anaphase I. Additionally, telomeres tethered at the nuclear envelope cluster in the bouquet configuration where they are subjected to dramatic pulling forces acting from outside of the nucleus. In Saccharomyces cerevisiae, the telomere-associated protein Ndj1 is required for bouquet formation. When Ndj1 is absent, these dramatic motions cease and multiple steps of meiosis I prophase progression are delayed. Here we identified Nup2 in a pool of enriched proteins that co-purify with tagged Ndj1 from meiotic cell extracts. Nup2 is a nonessential nucleoporin that functions in nuclear transport, boundary activity, and telomere silencing in mitotically dividing cells. We found that deletion of NUP2 delayed pairing and synapsis during meiosis, and led to decreased spore viability, similar to the ndj1{Delta} mutant phenotype. Surprisingly, the nup2{Delta} ndj1{Delta} double mutant failed to segregate chromosomes, even though the meiotic program continued. These results suggest that a physical impediment to nuclear division is created in the absence of Nup2 and Ndj1. Our deletion analysis of NUP2 identified a previously uncharacterized 125-amino acid region that is both necessary and sufficient to complement all of nup2{Delta}s meiotic phenotypes, which we call the meiotic autonomous region (MAR). We propose that Ndj1 and Nup2 function in parallel pathways to promote the dynamic chromosome events of meiotic chromosome dynamics, perhaps through the establishment or maintenance of higher-order chromosome organization.

Genetics

Nonessential nucleoporins Nup2, Nup60 and Nup84 are required for normal meiosis in budding yeast

The nuclear pore complex (NPC) selectively transports cargo between the nucleus and the cytoplasm. The inner nuclear membrane (INM) face of the NPC also serves as a hub where gene silencing and DNA repair are spatially coordinated. In Saccharomyces cerevisiae, partitioning of active and silenced chromatin at subtelomeric regions depends on the boundary activity of nonessential nucleoporin proteins Nup2 and Nup60, along with Htz1, the histone variant H2A.Z. The INM is also important for the chromosome events of meiosis since Ndj1-mediated telomere attachment and clustering at the INM is required for efficient homolog pairing, recombination, and segregation. Here we tested possible meiotic roles for Nup2, Nup60, Htz1, and other nonessential nucleoporins by analyzing the effects of deletion mutations on sporulation, spore viability and possible phenotypic epistasis with ndj1{Delta}. Deleting NUP2, NUP60, and HTZ1 reduced spore viability compared to wild-type (WT). A detailed analysis of spore lethality indicated that homolog nondisjunction in these mutants was elevated compared to WT, yet unlike ndj1{Delta}, this was not the predominant cause of spore death. Deleting NUP84 reduced meiosis I nuclear divisions, while deleting NUP53, NUP100, and NUP157 had no effect on sporulation, spore viability, or the kinetics of meiosis I progression. Surprisingly, nup2{Delta} ndj1{Delta} uniquely failed to undergo meiosis I nuclear divisions, suggesting Nup2 and Ndj1 function in partially redundant pathways or create a poisonous intermediate. The meiosis I division was also delayed by 2 hours in nup2{Delta} compared to WT pointing to a specialized role for Nup2 in the meiotic program.

Genetics

A computational approach to estimating nondisjunction frequency in Saccharomyces cerevisiae

Errors segregating homologous chromosomes during meiosis result in the formation of aneuploid gametes and are the largest contributing factor to birth defects and spontaneous abortions in humans. Saccharomyces cerevisiae has long served as a model organism for studying the gene network supporting normal chromosome segregation. Current methods of measuring homolog nondisjunction frequencies are laborious and involve dissecting thousands of tetrads to detect missegregation of individually marked chromosomes. Here we describe a holistic computational approach to determine the relative contributions of meiosis I nondisjunction and random spore death in mutants with reduced spore viability. These values are based on best-fit distributions of 4, 3, 2, 1, and 0 viable-spore tetrads to observed distributions in mutant and wild-type strains. We show proof-of-principle using published data sets that the calculated average meiosis I nondisjunction frequency closely matches empirically determined values. This analysis also points to meiosis I nondisjunction as an intrinsic component of spore inviability in wild-type strains. We uncover two classes of mutants that show distinct relationships between nondisjunction death and random spore death. Class I mutants, including those with known defects in establishing and maintaining the physical engagement of homologous chromosomes display a 4-fold greater ratio of nondisjunction death to random spore death compared to Class II mutants, which include those with defects in sister chromatid cohesion. Low numbers of required tetrads facilitates epistasis analysis to probe genetic interactions. Finally the application of the R-Scripts does not require any special strain construction and can be applied to previously observed tetrad distributions.

Genetics