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Dartmouth College

Recombinant fusion proteins

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Full-Text Articles in Molecular Biology

The Filament-Forming Protein Pil1 Assembles Linear Eisosomes In Fission Yeast, Ruth Kabeche, Suzanne Baldissard, John Hammond, Louisa Howard, James B. Moseley Aug 2011

The Filament-Forming Protein Pil1 Assembles Linear Eisosomes In Fission Yeast, Ruth Kabeche, Suzanne Baldissard, John Hammond, Louisa Howard, James B. Moseley

Dartmouth Scholarship

The cortical cytoskeleton mediates a range of cellular activities such as endocytosis, cell motility, and the maintenance of cell rigidity. Traditional polymers, including actin, microtubules, and septins, contribute to the cortical cytoskeleton, but additional filament systems may also exist. In yeast cells, cortical structures called eisosomes generate specialized domains termed MCCs to cluster specific proteins at sites of membrane invaginations. Here we show that the core eisosome protein Pil1 forms linear cortical filaments in fission yeast cells and that purified Pil1 assembles into filaments in vitro. In cells, Pil1 cortical filaments are excluded from regions of cell growth and are …


Requirements For Transitional Endoplasmic Reticulum Site Structure And Function In Saccharomyces Cerevisiae, Polina Shindiapina, Charles Barlowe Feb 2010

Requirements For Transitional Endoplasmic Reticulum Site Structure And Function In Saccharomyces Cerevisiae, Polina Shindiapina, Charles Barlowe

Dartmouth Scholarship

Secretory proteins are exported from the endoplasmic reticulum (ER) at specialized regions known as the transitional ER (tER). Coat protein complex II (COPII) proteins are enriched at tER sites, although the mechanisms underlying tER site assembly and maintenance are not understood. Here, we investigated the dynamic properties of tER sites in Saccharomyces cerevisiae and probed protein and lipid requirements for tER site structure and function. Thermosensitive sec12 and sec16 mutations caused a collapse of tER sites in a manner that depended on nascent secretory cargo. Continual fatty acid synthesis was required for ER export and for normal tER site structure, …


Meiotic Cohesion Requires Accumulation Of Ord On Chromosomes Before Condensation, Eric M. Balicky, Matthew W. Endres, Cary Lai, Sharon E. Bickel Sep 2002

Meiotic Cohesion Requires Accumulation Of Ord On Chromosomes Before Condensation, Eric M. Balicky, Matthew W. Endres, Cary Lai, Sharon E. Bickel

Dartmouth Scholarship

Cohesion between sister chromatids is a prerequisite for accurate chromosome segregation during mitosis and meiosis. To allow chromosome condensation during prophase, the connections that hold sister chromatids together must be maintained but still permit extensive chromatin compaction. In Drosophila, null mutations in the orientation disruptor (ord) gene lead to meiotic nondisjunction in males and females because cohesion is absent by the time that sister kinetochores make stable microtubule attachments. We provide evidence that ORD is concentrated within the extrachromosomal domains of the nuclei ofDrosophila primary spermatocytes during early G2, but accumulates on the meiotic chromosomes by …


Erv14p Directs A Transmembrane Secretory Protein Into Copii-Coated Transport Vesicles, Jacqueline Powers, Charles Barlowe Feb 2002

Erv14p Directs A Transmembrane Secretory Protein Into Copii-Coated Transport Vesicles, Jacqueline Powers, Charles Barlowe

Dartmouth Scholarship

Erv14p is a conserved integral membrane protein that traffics in COPII-coated vesicles and localizes to the early secretory pathway in yeast. Deletion of ERV14 causes a defect in polarized growth because Axl2p, a transmembrane secretory protein, accumulates in the endoplasmic reticulum and is not delivered to its site of function on the cell surface. Herein, we show that Erv14p is required for selection of Axl2p into COPII vesicles and for efficient formation of these vesicles. Erv14p binds to subunits of the COPII coat and binding depends on conserved residues in a cytoplasmically exposed loop domain of Erv14p. When mutations are …