Fusion between two eukaryotic cells is a fundamentally interesting and biologically important process. We have only a rudimentary understanding of how cell fusion occurs. This grant proposal focuses on characterizing the process of anastomosis (vegetative hyphal cell fusion) in the filamentous fungus Neurospora crassa. The major goals of the proposed research are to identify and characterize proteins that function to mediate the cell fusion event and to determine how they function to facilitate cell fusion. The genes that are required for cell fusion, as defined by cell fusion-defective mutants, will be identified and characterized. The mutations responsible for the cell fusion-defective phenotype will be mapped to small regions of the Neurospora genetic map by classical genetic mapping techniques. The affected genes will be identified using a PCR amplification and DNA sequencing strategy. The use of this positional cloning approach has been made possible by the publication of the Neurospora genomic DNA sequence. The identity of the genes encoding cell fusion proteins will be verified by gene disruption experiments. Anastomosis can be characterized as a series of steps leading to a cell fusion event. To help us determine which proteins function is each of these steps, differential interference contrast microscope and confocal microscope time-lapse video recording systems will be used to characterize the cell fusion mutants and elucidate which cell fusion steps are blocked within these mutants. We will also determine the location and follow the movement of a few key cell fusion proteins during cell fusion events. The cellular location of cell fusion proteins will be determined in immunolocalization experiments. Key cell fusion proteins will be tagged with the green fluorescent protein and the movement of these tagged proteins during cell fusion events will be followed with the confocal microscope time-lapse video recording system. We anticipate that these experiments will provide us with important insights into how cell fusion events are orchestrated. Public Health Relevance: Cell fusion events are critical to the process of fertilization and for the differentiation of muscle, bone, and placenta. The proposed research will help us better understand cell fusion events. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM078589-02
Application #
7291622
Study Section
Special Emphasis Panel (ZRG1-CB-G (02))
Program Officer
Deatherage, James F
Project Start
2006-09-25
Project End
2010-08-31
Budget Start
2007-09-01
Budget End
2008-08-31
Support Year
2
Fiscal Year
2007
Total Cost
$179,615
Indirect Cost
Name
State University of New York at Buffalo
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Fu, Ci; Tanaka, Asuma; Free, Stephen J (2014) Neurospora crassa 1,3-?-glucan synthase, AGS-1, is required for cell wall biosynthesis during macroconidia development. Microbiology 160:1618-27
Chinnici, Jennifer L; Fu, Ci; Caccamise, Lauren M et al. (2014) Neurospora crassa female development requires the PACC and other signal transduction pathways, transcription factors, chromatin remodeling, cell-to-cell fusion, and autophagy. PLoS One 9:e110603
Fu, Ci; Ao, Jie; Dettmann, Anne et al. (2014) Characterization of the Neurospora crassa cell fusion proteins, HAM-6, HAM-7, HAM-8, HAM-9, HAM-10, AMPH-1 and WHI-2. PLoS One 9:e107773
Maddi, Abhiram; Dettman, Anne; Fu, Ci et al. (2012) WSC-1 and HAM-7 are MAK-1 MAP kinase pathway sensors required for cell wall integrity and hyphal fusion in Neurospora crassa. PLoS One 7:e42374
Maddi, Abhiram; Fu, Ci; Free, Stephen J (2012) The Neurospora crassa dfg5 and dcw1 genes encode ?-1,6-mannanases that function in the incorporation of glycoproteins into the cell wall. PLoS One 7:e38872
Fu, Ci; Iyer, Priyadarshini; Herkal, Amrita et al. (2011) Identification and characterization of genes required for cell-to-cell fusion in Neurospora crassa. Eukaryot Cell 10:1100-9
Riquelme, Meritxell; Yarden, Oded; Bartnicki-Garcia, Salomon et al. (2011) Architecture and development of the Neurospora crassa hypha -- a model cell for polarized growth. Fungal Biol 115:446-74
Aldabbous, Mash'el Salman; Roca, M Gabriela; Stout, Angela et al. (2010) The ham-5, rcm-1 and rco-1 genes regulate hyphal fusion in Neurospora crassa. Microbiology 156:2621-9
Maddi, Abhiram; Bowman, Shaun M; Free, Stephen J (2009) Trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi Neurospora crassa and Candida albicans. Fungal Genet Biol 46:768-81
Bowman, Shaun M; Piwowar, Amy; Arnone, Eric D et al. (2009) Characterization of GPIT-1 and GPIT-2, two auxiliary components of the Neurospora crassa GPI transamidase complex. Mycologia 101:764-72

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