This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Deposition of the centromere-specific histone CENP-A (Cse4 in S. cerevisiae) is a key initial step in the formation of functional kinetochores. Histone deposition complex chromatin assembly factor I (CAF-I) plays a role in building centromedc chromatin. The CAC1, CAC2 and CAC3 genes (which encode the yeast CAF-I subunits) are not essential for viability, but deleting them decreases the stability of gene silencing at all heterochromatic loci, and cac mutants display altered nuclear morphology. Hir proteins have been implicated in nucleosome formation and heterochromatic gene silencing. Mutants lacking both of these genes (cacA hirA) display increased rates of chromosome missegregation and alterations in centromeric chromatin structure. These yeast show mislocalization of Cse4 (Sharp et al.) with indirect immunofluorescence on spread chromatin and multiple foci of Cse4-GFP fusion protein in live cells (Figure D.2.b.l.). We will use cryo x-ray tomography to: 1) examine the nuclear morphology of the single and double mutants to identify the abnormalities that result from deletion of these gene products; and 2) examine the distribution of Cse4 in the double mutant (cacb hirA) yeast.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR019664-04
Application #
7602925
Study Section
Special Emphasis Panel (ZRG1-F05 (40))
Project Start
2007-05-01
Project End
2008-04-30
Budget Start
2007-05-01
Budget End
2008-04-30
Support Year
4
Fiscal Year
2007
Total Cost
$10,368
Indirect Cost
Name
University of California San Francisco
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
McHugh, Emma; Batinovic, Steven; Hanssen, Eric et al. (2015) A repeat sequence domain of the ring-exported protein-1 of Plasmodium falciparum controls export machinery architecture and virulence protein trafficking. Mol Microbiol 98:1101-14
Do, Myan; Isaacson, Samuel A; McDermott, Gerry et al. (2015) Imaging and characterizing cells using tomography. Arch Biochem Biophys 581:111-21
Dasgupta, Sabyasachi; Auth, Thorsten; Gov, Nir S et al. (2014) Membrane-wrapping contributions to malaria parasite invasion of the human erythrocyte. Biophys J 107:43-54
Le Gros, Mark A; McDermott, Gerry; Cinquin, Bertrand P et al. (2014) Biological soft X-ray tomography on beamline 2.1 at the Advanced Light Source. J Synchrotron Radiat 21:1370-7
Smith, Elizabeth A; Cinquin, Bertrand P; Do, Myan et al. (2014) Correlative cryogenic tomography of cells using light and soft x-rays. Ultramicroscopy 143:33-40
Hanssen, Eric; Dekiwadia, Chaitali; Riglar, David T et al. (2013) Electron tomography of Plasmodium falciparum merozoites reveals core cellular events that underpin erythrocyte invasion. Cell Microbiol 15:1457-72
Smith, Elizabeth A; Cinquin, Bertrand P; McDermott, Gerry et al. (2013) Correlative microscopy methods that maximize specimen fidelity and data completeness, and improve molecular localization capabilities. J Struct Biol 184:12-20
Parkinson, Dilworth Y; Epperly, Lindsay R; McDermott, Gerry et al. (2013) Nanoimaging cells using soft X-ray tomography. Methods Mol Biol 950:457-81
Isaacson, Samuel A; Larabell, Carolyn A; Le Gros, Mark A et al. (2013) The influence of spatial variation in chromatin density determined by X-ray tomograms on the time to find DNA binding sites. Bull Math Biol 75:2093-117
McDermott, Gerry; Le Gros, Mark A; Larabell, Carolyn A (2012) Visualizing cell architecture and molecular location using soft x-ray tomography and correlated cryo-light microscopy. Annu Rev Phys Chem 63:225-39

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