The long-term objectives of this research are to understand the mechanism of bouquet formation using fission yeast as a model organism.
The specific aims are identification of regulatory and structural components involved in this process by genetic and biochemical strategies. For genetic study, screen for mutants defective in telomere clustering was conducted by observation of the dynamic distribution pattern of a heterochromatin binding protein - Swi6::GFP. Six mutants were obtained and found defining four genes. Two are known genes. Cloning of the remaining genes is undergoing and they will be further extensively characterized on the molecular level. For biochemical study, effort is being made to isolate and purify the telomere and SPB protein complex by tagged affinity purification and nucleus subfractionation. Components of the protein complex will be identified by mass-spectrometry and confirmed by localization of the in vivo tagged proteins. Since bouquet formation is a highly conserved process during meiotic prophase in most eukaryote and has been thought to facilitate the pairing of homologous chromosome, any knowledge of its mechanism will shed light on our understanding of male sterility, preterminated pregnancy and some genetic diseases.
| Jin, Ye; Mancuso, Joel J; Uzawa, Satoru et al. (2005) The fission yeast homolog of the human transcription factor EAP30 blocks meiotic spindle pole body amplification. Dev Cell 9:63-73 |
