The long-term goals of this project are to study the control and nature of recombination at the am locus of Neurospora, as well as the control of the expression of Glutamate dehydrogenase, the product of this gene. Glutamate dehydrogenase, an enzyme which is central in nitrogen metabolism of all eucaryotes, is highly conserved. The enzyme from Neurospora shows considerable homology, particularly in the amino terminal half, with that from vertebrates. Recent evidence has suggested that Glutamate dehydrogenase defects in humans are associated with a variety of neurological disorders including spinocerebellar syndrome. The difficulty of carrying out a program of fine structure genetic dissection of the gene of the glutamate dehydrogenase in higher eucaryotes makes this project with a microbial eucaryote particularly significant. As part of this current proposal the following studies will be carried out: (1) Recombination controlling elements in the 5 prime non-coding DNA will be identified and characterized (2) Mutations in the 5 prime non-coding DNA that effect gene expression will be characterized (3) The possibility of a distal element in the 5 prime non-coding DNA that effects gene expression will be examined (4) The nature of transformants obtained with the cloned am+ gene in a variety of vectors, using an am deletion as a recipient will be determined (5) The nature of non-homologous integration of am+ DNA during transformation will be determined by cloning and DNA sequencing the joint structure (6) Genes that flank the am gene will be cloned using the technique of chromosome walking.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM023967-10
Application #
3271985
Study Section
Genetics Study Section (GEN)
Project Start
1978-09-01
Project End
1990-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
10
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Kansas
Department
Type
Schools of Medicine
DUNS #
016060860
City
Kansas City
State
KS
Country
United States
Zip Code
66160
Perkins, D D; Kinsey, J A; Asch, D K et al. (1993) Chromosome rearrangements recovered following transformation of Neurospora crassa. Genetics 134:729-36
Asch, D K; Frederick, G; Kinsey, J A et al. (1992) Analysis of junction sequences resulting from integration at nonhomologous loci in Neurospora crassa. Genetics 130:737-48
Foss, E J; Garrett, P W; Kinsey, J A et al. (1991) Specificity of repeat-induced point mutation (RIP) in Neurospora: sensitivity of non-Neurospora sequences, a natural diverged tandem duplication, and unique DNA adjacent to a duplicated region. Genetics 127:711-7
Frederick, G D; Kinsey, J A (1990) Nucleotide sequence and nuclear protein binding of the two regulatory sequences upstream of the am (GDH) gene in Neurospora. Mol Gen Genet 221:148-54
Kinsey, J A (1990) Tad, a LINE-like transposable element of Neurospora, can transpose between nuclei in heterokaryons. Genetics 126:317-23
Frederick, G D; Kinsey, J A (1990) Distant upstream regulatory sequences control the level of expression of the am (GDH) locus of Neurospora crassa. Curr Genet 18:53-8
Asch, D K; Kinsey, J A (1990) Relationship of vector insert size to homologous integration during transformation of Neurospora crassa with the cloned am (GDH) gene. Mol Gen Genet 221:37-43
Kinsey, J A; Helber, J (1989) Isolation of a transposable element from Neurospora crassa. Proc Natl Acad Sci U S A 86:1929-33
Kinsey, J A (1989) Restricted distribution of the Tad transposon in strains of Neurospora. Curr Genet 15:271-5
Frederick, G D; Asch, D K; Kinsey, J A (1989) Use of transformation to make targeted sequence alterations at the am (GDH) locus of Neurospora. Mol Gen Genet 217:294-300