scholarly journals The vacuolar ATPase of Neurospora crassa contains an F1-like structure

1989 ◽  
Vol 264 (26) ◽  
pp. 15606-15612
Author(s):  
B J Bowman ◽  
W J Dschida ◽  
T Harris ◽  
E J Bowman
Keyword(s):  
Neurospora Crassa ◽  
Vacuolar Atpase

Vacuolar ATPase of Neurospora crassa: electron microscopy, gene characterization and gene inactivation/mutation.

1992 ◽  
Vol 172 (1) ◽  
pp. 57-66
Author(s):  
B J Bowman ◽  
W J Dschida ◽  
E J Bowman
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Neurospora Crassa ◽  
Specific Inhibitor ◽  
Vacuolar Atpase ◽  
Gene Inactivation ◽  
Bafilomycin A1 ◽  
Gene Characterization ◽  
Atpase Gene ◽  
Genes Encoding

We are using three approaches to investigate the vacuolar ATPase, V-ATPase, from Neurospora crassa. (1) Examination in the electron microscope shows the enzyme has a 'ball and stalk' structure like the F-type ATPases. However, the vacuolar ATPase is significantly larger, has a prominent cleft in the head sector, and has extra components associated with the stalk and membrane sectors. (2) Genes encoding three of the major subunits of the vacuolar ATPase and the homologous subunits of the mitochondrial F-ATPase have been isolated. The exon/intron structures of the genes have been analyzed and the chromosomal locations have been determined. Two of the vacuolar ATPase genes map very close to each other, suggesting the possibility of a cluster of ATPase genes. (3) The function of the ATPase is being investigated by isolating strains with altered or inactivated ATPase. We are characterizing strains that are resistant to bafilomycin A1, a potent and specific inhibitor of the vacuolar ATPase. Initial attempts to inactivate a vacuolar ATPase gene indicate that the enzyme may be essential for growth.

scholarly journals A Model for the Proteolipid Ring and Bafilomycin/Concanamycin-binding Site in the Vacuolar ATPase of Neurospora crassa

2006 ◽  
Vol 281 (42) ◽  
pp. 31885-31893
Author(s):  
Barry J. Bowman ◽  
Mary E. McCall ◽  
Robert Baertsch ◽  
Emma Jean Bowman
Keyword(s):  
Neurospora Crassa ◽  
Binding Site ◽  
Vacuolar Atpase

The Vacuolar ATPase of Neurospora crassa Is Indispensable: Inactivation of the vma-1 Gene by Repeat-Induced Point Mutation

Genetics ◽  
1996 ◽  
Vol 143 (1) ◽  
pp. 147-154 ◽  
Author(s):  
Tracy L Ferea ◽  
Barry J Bowman
Keyword(s):  
Neurospora Crassa ◽  
Point Mutation ◽  
Point Mutations ◽  
Vacuolar Atpase ◽  
Catalytic Subunit ◽  
Multiple Point ◽  
Extra Copy ◽  
Process Sequencing ◽  
Functional Copy ◽  
Repeat Induced Point Mutation

Abstract To analyze the phenotype of cells lacking the vacuolar ATPase, we inactivated the vma-1 gene, which encodes the catalytic subunit of the enzyme. Because preliminary experiments suggested the vma-1 gene was essential, we developed a method of simultaneously inactivating the gene and complementing it with a functional copy. We call this method repeat-induced point mutation (RIP) & Rescue. Two strains, both of which contained an extra copy of the vma-1 gene, were mated. Progeny that had inherited a functional copy of the gene at an ectopic site in the genome were selected. In some of these progeny the endogenous vma-1 gene had been altered by the RIP process. Sequencing showed the endogenous vma-1 gene had been inactivated by multiple point mutations. Progeny from strains with an inactive endogenous vma-1 gene were inviable unless a functional copy of the gene cosegregated, indicating that the vacuolar ATPase is essential in Neurospora crassa.

Ultrastructure and Morphology of the Neurospora Crassa Cell Wall Mutants, Slime And Slime X, Using Tem and Sem

1976 ◽  
Vol 34 ◽  
pp. 54-55
Author(s):  
Karen S. Howard ◽  
H. D. Braymer ◽  
M. D. Socolofsky ◽  
S. A. Milligan
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Wild Type ◽  
Morphological Comparison ◽  
Small Areas ◽  
Solid Media ◽  
Thin Sectioning ◽  
X Cells ◽  
Mutant Cells ◽  
Isolated Cell

The recently isolated cell wall mutant slime X of Neurospora crassa was prepared for ultrastructural and morphological comparison with the cell wall mutant slime. The purpose of this article is to discuss the methods of preparation for TEM and SEM observations, as well as to make a preliminary comparison of the two mutants.TEM: Cells of the slime mutant were prepared for thin sectioning by the method of Bigger, et al. Slime X cells were prepared in the same manner with the following two exceptions: the cells were embedded in 3% agar prior to fixation and the buffered solutions contained 5% sucrose throughout the procedure.SEM: Two methods were used to prepare mutant and wild type Neurospora for the SEM. First, single colonies of mutant cells and small areas of wild type hyphae were cut from solid media and fixed with OSO4 vapors similar to the procedure used by Harris, et al. with one alteration. The cell-containing agar blocks were dehydrated by immersion in 2,2-dimethoxypropane (DMP).

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