Functional analyses of the Neurospora crassa MT a-1 mating type polypeptide.
The Neurospora crassa mt a-1 gene, encoding the MT a-1 polypeptide, determines a mating type properties: sexual compatibility and vegetative incompatibility with A mating type. We characterized in vivo and in vitro functions of the MT a-1 polypeptide and specific mutant derivatives. MT a-1 polypeptide produced in Escherichia coli bound to specific DNA sequences whose core was 5'-CTTTG-3'. DNA binding was a function of the MT a-1 HMG box domain (a DNA binding motif found in high mobility group proteins and a diverse set of regulatory proteins). Mutation within the HMG box eliminated DNA binding in vitro and eliminated mating in vivo, but did not interfere with vegetative incompatibility function in vivo. Conversely, deletion of amino acids 216-220 of MT a-1 eliminated vegetative incompatibility, but did not affect mating or DNA binding. Deletion of the carboxyl terminal half of MT a-1 eliminated both mating and vegetative incompatibility in vivo, but not DNA binding in vitro. These results suggest that mating depends upon the ability of MT a-1 polypeptide to bind to, and presumably to regulate the activity of, specific DNA sequences. However, the separation of vegetative incompatibility from both mating and DNA binding indicates that vegetative incompatibility functions by a biochemically distinct mechanism.
Philley
M L
ML
T. H. Morgan School of Biological Sciences, University of Kentucky, Lexington 40506-0225.
Staben
C
C
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D002459:Cell Fusion; D003433:Crosses, Genetic; D004271:DNA, Fungal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D049770:Genes, Mating Type, Fungal; D006609:High Mobility Group Proteins / Q000737:chemistry* / Q000502:physiology; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D011487:Protein Conformation; D012100:Reproduction, Asexual; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013172:Spores, Fungal / Q000502:physiology; D013329:Structure-Activity Relationship
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D006609:High Mobility Group Proteins