Structure and Distribution of Organelles and Cellular Location of Calcium Transporters in Neurospora crassa

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Published inEukaryotic Cell Vol. 8; no. 12; pp. 1845 - 1855
Main Authors Bowman, Barry J, Draskovic, Marija, Freitag, Michael, Bowman, Emma Jean
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.12.2009
American Society for Microbiology (ASM)
Subjects
Adenosine Triphosphatases - metabolism
Biomarkers - 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
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Abstract Classifications Services EC Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue EC About EC Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy EC RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1535-9778 Online ISSN: 1535-9786 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to EC .asm.org, visit: EC       
AbstractList 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 μm 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.
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.
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 μm 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.
Classifications Services EC Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue EC About EC Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy EC RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1535-9778 Online ISSN: 1535-9786 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to EC .asm.org, visit: EC       
Author Marija Draskovic
Michael Freitag
Barry J. Bowman
Emma Jean Bowman
AuthorAffiliation Department of Molecular, Cell, and Developmental Biology, University of California; Santa Cruz, California 95064, 1 Department of Biochemistry and Biophysics, Oregon State University, ALS 2011, Corvallis, Oregon 97331-7305 2
AuthorAffiliation_xml – name: Department of Molecular, Cell, and Developmental Biology, University of California; Santa Cruz, California 95064, 1 Department of Biochemistry and Biophysics, Oregon State University, ALS 2011, Corvallis, Oregon 97331-7305 2
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  surname: Bowman
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  organization: Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, CA 95064, USA. bowman@biology.ucsc.edu
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  fullname: Freitag, Michael
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  givenname: Emma Jean
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/19801418$$D View this record in MEDLINE/PubMed
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We wanted to examine the cellular locations of four Neurospora crassa proteins that transport calcium. However, the structure and distribution of organelles in...
ABSTRACT We wanted to examine the cellular locations of four Neurospora crassa proteins that transport calcium. However, the structure and distribution of...
We wanted to examine the cellular locations of four Neurospora crassa proteins that transport calcium. However, the structure and distribution of organelles in...
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StartPage 1845
SubjectTerms Adenosine Triphosphatases - metabolism
Biomarkers - 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
Title Structure and Distribution of Organelles and Cellular Location of Calcium Transporters in Neurospora crassa
URI http://ec.asm.org/content/8/12/1845.abstract
https://www.ncbi.nlm.nih.gov/pubmed/19801418
https://search.proquest.com/docview/733130301
https://pubmed.ncbi.nlm.nih.gov/PMC2794220
Volume 8
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