X-ray-induced specific locus mutations in the ad-3 region of two-component heterokaryons of Neurospora crassa. I. Modification of the heterozygous effects of multilocus deletions covering the ad-3A or ad-3B loci.

The basis for the reduced growth rates of heterokaryons between strains carrying nonallelic combinations of gene/point mutations (ad-3R) and multilocus deletion mutations (ad-3IR) has been investigated by a simple genetic test. The growth rates of forced 2-component heterokaryons (dikaryons) between multilocus deletion mutations were compared with forced 3-component heterokaryons (trikaryons) containing an ad-3AR ad-3BR double mutant as their third component. Since the third component has no genetic damage at other loci immediately adjacent to the ad-3A or ad-3B locus, the growth rate on minimal medium depends on the functional activity of the unaltered (and presumed "wild-type") ad-3A and ad-3B loci in the first two components. In many cases, the requirements of the original dikaryons have been satisfied by the addition of unaltered genes (in the third component), and these trikaryons grow at wild-type rate on minimal medium. Those trikaryons growing at less than wild-type rate were shown to be adenine-requiring, and wild-type growth rate was obtained with the addition of low levels of adenine to the medium. Such tests in the present experiments have shown that ad-3IR mutations result not only in inactivation of the ad-3 loci by multilocus deletion but also, in many cases, in partial gene inactivation by an unknown mechanisms at other loci in the immediately adjacent regions. The heterozygous effects observed in our present experiments with multilocus deletions in Neurospora can be explained either by a spreading-type position effect of the type found by others in Drosophila, mice, Oenothera and Aspergillus or by undetected genetic damage ("cryptic mutations") in the immediately adjacent genetic regions. An attempt will be made to distinguish between these two alternative hypotheses with techniques for DNA cloning and sequencing in future experiments.