RESUMEN
The yeast Zip1 protein (Zip1p) is a component of the central region of the synaptonemal complex (SC). Zip1p is predicted to form a dimer consisting of a coiled-coil domain flanked by globular domains. To analyze the organization of Zip1p within the SC, in-frame deletions of ZIP1 were constructed and analyzed. The results demonstrate that the C terminus but not the N terminus of Zip1p is required for its localization to chromosomes. Deletions in the carboxy half of the predicted coiled-coil region cause decreases in the width of the SC. Based on these results, a model for the organization of Zip1p within the SC is proposed. zip1 deletion mutations were also examined for their effects on sporulation, spore viability, crossing over, and crossover interference. The results demonstrate that the extent of synapsis is positively correlated with the levels of spore viability, crossing over, and crossover interference. In contrast, the role of Zip1p in synapsis is separable from its role in meiotic cell cycle progression. zip1 mutants display interval-specific effects on crossing over.
Asunto(s)
Cromosomas Fúngicos/genética , Proteínas Fúngicas/genética , Meiosis/genética , Mutagénesis Sitio-Dirigida , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Anticuerpos Antifúngicos , Cromosomas Fúngicos/química , Cromosomas Fúngicos/inmunología , Cromosomas Fúngicos/metabolismo , Intercambio Genético/genética , Proteínas Fúngicas/inmunología , Proteínas Fúngicas/metabolismo , Proteínas Nucleares , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Sistemas de Lectura/genética , Esporas Fúngicas/genética , Coloración y Etiquetado , Complejo Sinaptonémico/genéticaRESUMEN
In continuation of our efforts to elucidate the immunoglobulin kappa locus of the mouse we analyzed 46 yeast artificial chromosomes (YACs) containing V kappa, J kappa and C kappa genes. The YACs, which were derived from DNA of C57BL/6 and C3H mice, ranged from 0.3-1.9 Mb in size. On the basis of hybridization with probes specific for the V kappa gene families a group of 13 YACs was selected for detailed analysis. The V kappa genes of the YACs were then characterized by hybridization to the family-specific probes and by the sizes of the EcoRI fragments on which they were found. This way evidence was obtained for 140 different V kappa gene signals on the YACs. Of these 63 had been characterized before on clones from a cosmid library of total mouse DNA (I. Zocher et al., Eur. J. Immunol. 1995. 25: 3326-3331) and 22 others were found now on cosmid clones derived from the YACs. Six V kappa genes of the previous study which were not found on the YACs are probably located outside of the kappa locus. The YACs were arrayed in a unique order establishing a YACs panel which most likely contains the whole kappa locus. The cosmid contigs and solitary cosmid clones which contain the 63 plus 22 V kappa gene signals mentioned above comprise about 2.0 Mb. Assuming that the remaining 55 V kappa genes are spaced at the same average distance of 24 kb, one may extrapolate to a locus size of 3.3 Mb.