Magnification in Drosophila: evidence for an inducible rDNA-specific recombination system.

An experiment is described that provides evidence for an exchange mechanism to explain the increase in ribosomal gene number that occurs during bobbed magnification. We show that bobbed and bobbed-lethal alleles do not magnify in closed X chromosomes, but that a spontaneous ring opening restores normal magnification. The results provide strong evidence that the elementary magnifying event is unequal sister chromatid exchange, and can be interpreted in the framework of an inducible rDNA-specific recombination system as the basis of ribosomal gene magnification.

Ring chromosomes and rDNA magnification in Drosophila.

Tartof showed that ribosomal gene magnification in Drosophila was inhibited in a ring X chromosome. The present studies extend this observation by showing that ring X chromosomes are lost meiotically in male Drosophila undergoing ribosomal gene magnification as evidenced by the recovery of a lower number of ring-bearing progeny under magnifying conditions compared with nonmagnifying conditions. Associated with ring chromosome loss is a highly significant increase in the number of double-sized dicentric ring chromosomes in meiotic cells from magnifying males. These observations explain the failure of ring X chromosomes to magnify and imply that magnification in rod chromosomes occurs via a mechanism of unequal sister chromatid exchange. Our results support the hypothesis that the primary event of magnification is a sister chromatid exchange in the rDNA, that the frequency of sister strand exchanges is increased in magnifying flies, that a significant number of exchanges in magnifying flies occurs meiotically and that some of the exchanges are nonreciprocal. We have also found that autosomal mutations can affect both the frequency of abnormal ring structures and the ability of ring X chromosomes to magnify.

The localization of mouse globin genes: a test of the effectiveness of hybridization in situ.

Hybridization of rabbit reticulocyte mRNA to banded mouse chromosomes in situ labeled several regions, including the globin loci. Whereas the labeling was sufficient to detect unknowns in the globin size class, the chromosome assignments would be doubtful without some means of removing trace contaminants from the probe or of recognizing chromosomal regions to which they hybridize. Mammalian gene mapping by means of hybridization in situ might be feasible with probes cloned in microbial host-vector systems or with kinetic analysis of the hybridization process at every labeled site.

The chromosomal location of rDNA in selected lower primates.

Hybridization in stiu was used to identify the chromosomes that carry rDNA in representative lower primates, including the baboons, Papio cynocephalus and Papio hamadryas; the colobus monkey, Colobus polykomos; the tree shrew, Tupaia glis; the lemur, Lemur fulvis; the saki, Pithecia pithecia; the marmoset, Saguinus nigricollis, and the spider monkey, Ateles geoffroyi. The marker chromosome, common to the Cercopithecines studied to date, carries the rDNA in the baboons. Another marker chromosome carries rDNA in a South American species, the spider monkey. A multichromosomal distribution of rDNA was demonstrated in the tree shrew, lemur, saki, and marmoset. None of the rDNA-containing chromosomes in the prosimians and New World monkeys show homology to the chromosomes that carry rDNA in the Hominids, Pongids, or Old World monkeys.

Chromosomal distribution of rDNA in Pan paniscus, Gorilla gorilla beringei, and Symphalangus syndactylus: comparison to related primates.

Hybridization in situ was used to identify rDNA in chromosomes of the pygmy chimpanzee, mountain gorilla, and siamang gibbon. In contrast to other Pongids, and man, the gorilla has only two pairs of rDNA-containing chromosomes. The single pair in the siamang bears no resemblance to the nucleolar chromosome of the closely related lar gibbon. Pan paniscus and P. troglodytes have the same rDNA distribution, and similar karyotypes except in the structure of chromosome 23p. Grain counts over unbanded preparations show that the human, orangutan, and both chimpanzees have about the same total rDNA multiplicity.

The site of 5S RNA genes in primates. I. The great apes.

A major site of genes for 5S RNA has been localized in representative members of the family Pongidae by means of hybridization in situ. These genes are shown to be concentrated in the most distal bands of the primate chromosome arm homologous to human chromosome 1q.

Structure and function of initiation complexes which accumulate during inhibition of protein synthesis by fluoride ion.

Incubation of the reticulocyte lysate cell-free system with KF results in the accumulation in polysomes of complexes containing deacylated tRNAMet and of complexes which can initiate globin chains in the presence of aurintricarboxylate. Degradation of these polysomes with T1RNase yields both 40 S and 80 S particles, and tRNAMet is found in both of these fractions. When the 80 S particles are reincubated with the soluble fraction of the lysate plus reagents for protein synthesis, short peptides which have the properties of the NH2-terminal regions of globin are synthesized de novo. These peptides are deficient in NH2-terminal methionine, but occur under conditions where nascent globin peptides of comparable length, containing NH2-terminal methionine, are completely protected from the methionine aminopeptidase.

Variation in the number of genes for rRNA among human acrocentric chromosomes: correlation with frequency of satellite association.

Grain counts after hybridization of 125I-rRNA to human chromosomes indicate numerical polymorphism at the rDNA sites. Prephotographing procedures decrease labeling but do not change the proportions of labeled RNA annealed to different chromosomes. A positive correlation was found between the frequency of participation of a given chromosome in satellite associations and its rDNA content by the criterion of grain count. Certain individual chromosomes are clear exceptions to this correlation.

Variation in ribosomal RNA gene number in mouse chromosomes.

Hybridization of 125-I-ribosomal RNA to mouse chromosomes in situ produced significant differences in grain count at known rDNA sites, depending on the strains from which they were derived. This is interpreted to mean that the number of rRNA genes in a given nucleolar chromosome, and in the entire genome, is polymorphic among strains and among outbred individuals.

Does the T-locus in the mouse include ribosomal DNA?

The proportion of rDNA in the mouse genome is shown to have no systematic relation to the T-locus genotype, and hybridization in situ with 125I-RNA does not label chromosome 17, which carries the T-locus. Thus rDNA need no longer be taken into account in explaining the unique properties of the alleles at this locus. The number of genes for rRNA precursor averaged 153 per haploid genome, but a large measure of individual variation is suggested.

Satellite-association frequency and rDNA content of a double-satellited chromosome.

A correlation between the amount of rDNA and the frequency of participation in satellite associations is observed in a double-satellited human acrocentric chromosome.