RESUMO
map has been developed of nuclease-hypersensitive sites of P-rr, the standard allele of the P-locus of Zea mays L. Using a traditional DNase I assay, eight such sites have been found that are specific for the expressing tissue and span a region of more than 25 kb of the P-locus, making it one of the largest plant genes yet described. The maps of the standard allele have also been compared with the recently described moderately stable P-pr allele, which arose from epimutation. Six of the eight sites exhibit the same tissue-specificity in P-pr plants, while two stay repressed as in non-expressing tissues of plants with the standard allele. Interestingly, the two repressed sites coincide with two hypermethylated restriction sites that have previously been correlated with the expression potential of the P-pr allele. On the other hand, four of the DNase I sites, coinciding with CpG islands that were not hypermethylated by the epimutation, also showed no differences in their sensitivity to DNase I between the standard allele and the P-pr allele. This suggests that the epimutation affects both site-specific methylation changes and a specific local chromatin structure of the P gene involved in its regulation.
RESUMO
map has been developed of nuclease-hypersensitive sites of P-rr, the standard allele of the P-locus of Zea mays L. Using a traditional DNase I assay, eight such sites have been found that are specific for the expressing tissue and span a region of more than 25 kb of the P-locus, making it one of the largest plant genes yet described. The maps of the standard allele have also been compared with the recently described moderately stable P-pr allele, which arose from epimutation. Six of the eight sites exhibit the same tissue-specificity in P-pr plants, while two stay repressed as in non-expressing tissues of plants with the standard allele. Interestingly, the two repressed sites coincide with two hypermethylated restriction sites that have previously been correlated with the expression potential of the P-pr allele. On the other hand, four of the DNase I sites, coinciding with CpG islands that were not hypermethylated by the epimutation, also showed no differences in their sensitivity to DNase I between the standard allele and the P-pr allele. This suggests that the epimutation affects both site-specific methylation changes and a specific local chromatin structure of the P gene involved in its regulation.
RESUMO
Developmental changes in the plasma membrane proteins of Dictyostelium discoideum have been studied using metabolic labeling with [35S]methionine and two-dimensional electrophoresis. Pulse labeling for 1 h at the early interphase, late interphase, aggregation, and tip formation stages of development showed that the profile of newly synthesized plasma membrane proteins changed dramatically over this interval. Only 14% of the polypeptide species were synthesized at all four stages at detectable levels; 86% of the species changed over this developmental interval according to the criterion that they were synthesized at some but not all of the four stages tested. Long-term labeling during vegetative growth followed by initiation of development showed that the "steady-state" levels of the plasma membrane proteins changed very little over the same period. The only changes were in minor species (33% overall change). Similar analyses of whole cell proteins showed 27 and 20% change, respectively. Cell surface radioiodination revealed 52 external proteins in the plasma membrane. Comparison with the uniform methionine labeling results showed that these proteins were, with one notable exception, minor membrane components. In these external proteins, also, developmental changes were limited and were observed in the less abundant species. These results demonstrate the existence of two general classes of plasma membrane proteins. The first is a population of high-abundance proteins that are present in vegetative cells and are largely conserved through development. These possibly serve "housekeeping" functions common to all stages. The second class consists of low-abundance species that are expressed in a highly stage-specific manner and which presumably participate in developmentally important functions.
