PMID:19801418

Citation	Bowman, BJ, Draskovic, M, Freitag, M and Bowman, EJ (2009) Structure and distribution of organelles and cellular location of calcium transporters in Neurospora crassa. Eukaryotic Cell 8:1845-55
Abstract	We wanted to examine the cellular locations of four Neurospora crassa proteins that transport calcium. However, the structure and distribution of organelles in live hyphae of N. crassa have not been comprehensively described. Therefore, we made recombinant genes that generate translational fusions of putative organellar marker proteins with green or red fluorescent protein. We observed putative endoplasmic reticulum proteins, encoded by grp-78 and dpm, in the nuclear envelope and associated membranes. Proteins of the vacuolar membrane, encoded by vam-3 and vma-1, were in an interconnected network of small tubules and vesicles near the hyphal tip, while in more distal regions they were in large and small spherical vacuoles. Mitochondria, visualized with tagged ARG-4, were abundant in all regions of the hyphae. Similarly, we tagged the four N. crassa proteins that transport calcium with green or red fluorescent protein to examine their cellular locations. NCA-1 protein, a homolog of the SERCA-type Ca(2+)-ATPase of animal cells, colocalized with the endoplasmic reticulum markers. The NCA-2 and NCA-3 proteins are homologs of Ca(2+)-ATPases in the vacuolar membrane in yeast or in the plasma membrane in animal cells. They colocalized with markers in the vacuolar membrane, and they also occurred in the plasma membrane in regions of the hyphae more than 1 mm from the tip. The cax gene encodes a Ca(2+)/H(+) exchange protein found in vacuoles. As expected, the CAX protein localized to the vacuolar compartment. We observed, approximately 50 to 100 mum from the tip, a few spherical organelles that had high amounts of tagged CAX protein and tagged subunits of the vacuolar ATPase (VMA-1 and VMA-5). We suggest that this organelle, not described previously in N. crassa, may have a role in sequestering calcium.
Links	PubMed PMC2794220 Online version:10.1128/EC.00174-09
Keywords	Adenosine Triphosphatases/metabolism; Biological Markers/metabolism; Calcium/metabolism; Cell Membrane/metabolism; Endoplasmic Reticulum/metabolism; Fungal Proteins/metabolism; Golgi Apparatus/metabolism; Hyphae/metabolism; Membrane Transport Proteins/metabolism; Mitochondria/metabolism; Models, Biological; Neurospora crassa/cytology; Neurospora crassa/metabolism; Nuclear Envelope/metabolism; Organelles/metabolism; Protein Transport; Vacuoles/enzymology
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
NEUCS:Q9UUY2		GO:0005773: vacuole	ECO:0000314:		C		Figure 4C shows nca2 colocalizes with the vacuole marker protein vam3.	complete CACAO 6035
NEUCS:Q9UUY2	part_of	GO:0005773: vacuole	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
NEUCS:Q9UUY0		GO:0005635: nuclear envelope	ECO:0000314:		C		Figure 2C shows nca-1 is located in the nuclear envelope; the nucleus is marked with RFP::hpo.	complete CACAO 6037
NEUCS:Q9UUY0	part_of	GO:0005635: nuclear envelope	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete

References

See Help:References for how to manage references in GONUTS.