Dual Sets of Chimeric Alleles Identify Specificity Sequences for the bE and bW Mating and Pathogenicity Genes of Ustilago maydis

ABSTRACT

The b mating-type locus of the fungal plant pathogenUstilago maydis encodes two multiallelic gene products, bE and bW, that control the formation and maintenance of the infectious cell type. Dimerization via the N-terminal regions of bE and bW proteins encoded by alleles of different specificities establishes a homeodomain-containing transcription factor. The bE and bW products encoded by alleles of like specificities fail to dimerize. We constructed sets of chimeric alleles for the bE1 and bE2 genes and for the bW1 and bW2genes to identify sequences that control specificity. The mating behavior of strains carrying chimeric alleles identified three classes of specificity: b2 (class I), specificity different from either parental type (class II), and b1 (class III). Crosses between strains carrying bE and bWchimeric alleles identified two short blocks of amino acids that influence specificity and that are located in the N-terminal variable regions of the b proteins. Comparisons of pairs of chimeric alleles encoding polypeptides differing in specificity and differing at single amino acid positions identified 16 codon positions that influence the interaction between bE and bW. Fifteen of these positions lie within the blocks of amino acids identified by crosses between the strains carrying chimeric alleles. Overall, this work provides insight into the organization of the regions that control recognition.

ACKNOWLEDGMENTS

This work was supported by Research and Collaborative Project grants from NSERC (to J.W.K.) and by postgraduate fellowships from NSERC and the Killam Foundation (to A.R.Y.).

We thank Jeanette Johnson-Beatty, George Athwal, and Alan Au for technical assistance and Lisa Gentile and Lawrence McIntosh for comments on the manuscript.