Search bioRxiv⌕ Search

Biology subjects

Bestul, A. J.

Publications and source records attributed to Bestul, A. J..

3 recordsLinked to original sources

Fss1, a CTNS/PQ domain protein, has dual roles in promoting fertility in Schizosaccharomyces pombe

Sexual reproduction is critical in most eukaryotes, but many genes required for the process remain to be identified and characterized. A previous saturation mutagenesis study in Schizosaccharomyces pombe implicated several unnamed genes in having roles in sexual reproduction. In this work, we provide an initial characterization of 13 of these genes and an in-depth characterization of one gene, fss1 (facilitates starvation sex). Fss1 has high structural homology to the human PQ-loop repeat family PQLC2 transceptor (amino acid transporter plus membrane receptor), but likely lacks the ability to transport amino acids due to loss of a critical PQ motif. fss1{Delta} mutants have no defects in meiosis but have pronounced defects in packaging the products of meiosis into spores, particularly in nutrient-poor environments. The spore formation phenotypes are recessive as fss1{Delta} spores produced by a fss1+/fss1{Delta} heterozygote develop normally. Immuno-gold electron microscopy shows Fss1 localizes to vesicles that fuse to the ephemeral outer spore membrane as the spore wall matures below. Fss1 protein is also produced within spores and this population is required for normal vacuole morphology and timely germination. We propose a model in which Fss1 is a vesicle receptor that facilitates spore wall formation from diploid-expressed protein, while spore-expressed protein acts at the vacuole to promote an internal cellular environment that facilitates germination. This work expands our catalog of genes that contribute to sexual reproduction and identifies two previously unknown tasks in spore biology that are both facilitated by Fss1.

genetics↗

Quantitative analysis of nuclear pore complex organization in Schizosaccharomyces pombe

The number, distribution and composition of nuclear pore complexes (NPCs) in the nuclear envelope (NE) varies between cell types and changes during cellular differentiation and in disease. To understand how NPC density and organization is controlled, we analyzed NPC number and distribution in the fission yeast Schizosaccharomyces pombe using structured illumination microscopy. The small size of yeast nuclei, genetic features of fungi and our robust image analysis pipeline allowed us to study NPCs in intact nuclei under multiple conditions. Our data revealed that NPC density is maintained across a wide range of nuclear sizes. Regions of reduced NPC density are observed over the nucleolus and surrounding the spindle pole body (SPB). Lem2-mediated tethering of the centromeres to the SPB is required to maintain NPC exclusion, which is important for timely mitotic progression. These findings provide a quantitative understanding of NPC number and distribution in S. pombe and show that interactions between the centromere and the NE influences local NPC distribution.

cell biology↗

Redistribution of centrosomal proteins by centromeres and Polo kinase controls nuclear envelope breakdown

Proper mitotic progression in Schizosaccharomyces pombe requires partial nuclear envelope breakdown (NEBD) and insertion of the spindle pole body (SPB - yeast centrosome) to build the mitotic spindle. Linkage of the centromere to the SPB is vital to this process, but why that linkage is important is not well understood. Utilizing high- resolution structured illumination microscopy (SIM), we show that the conserved SUN- domain protein Sad1 and other SPB proteins redistribute during mitosis to form a ring complex around SPBs, which is a precursor for localized NEBD and spindle formation. Although the Polo kinase Plo1 is not necessary for Sad1 redistribution, it localizes to the SPB region connected to the centromere, and its activity is vital for redistribution of other SPB ring proteins and for complete NEBD at the SPB to allow for SPB insertion. Our results lead to a model in which centromere linkage to the SPB drives redistribution of Sad1 and Plo1 activation that in turn facilitate partial NEBD and spindle formation through building of a SPB ring structure.

cell biology↗