Ulrich Grossbach

Histone H1 Genes and Histone Gene Clusters in the Genus Drosophila

Abstract. Whereas the genomes of many organisms contain several nonallelic types of linker histone genes, one single histone H1 type is known in Drosophila melanogaster that occurs in about 100 copies per genome. Amplification of H1 gene sequences from genomic DNA of wild type strains of D. melanogaster from Oregon, Australia, and central Africa yielded numerous clones that all exhibited restriction patterns identical to each other and to those of the known H1 gene sequence. Nucleotide sequences encoding the evolutionarily variable domains of H1 were determined in two gene copies of strain Niamey from central Africa and were found to be identical to the known H1 sequence. Most likely therefore, the translated sequences of D. melanogaster H1 genes do not exhibit intragenomic or intergenomic variations.In contrast, three different histone H1 genes were isolated from D. virilis and found to encode proteins that differ remarkably from each other and from the H1 of D. melanogaster and D. hydei. About 40 copies of H1 genes are organized in the D. virilis genome with copies of core histone genes in gene quintets that were found to be located in band 25F of chromosome 2. Another type of histone gene cluster is present in about 15 copies per genome and contains a variable intergenic sequence instead of an H1 gene. The H1 heterogeneity in D. virilis may have arisen from higher recombination rates than occur near the H1 locus in D. melanogaster and might provide a basis for formation of different chromatin subtypes.

Localization of nuclear proteins related to high mobility group protein 14 (HMG 14) in polytene chromosomes

An antibody was raised against “high mobility group” nuclear protein 14 (HMG 14) from calf thymus, known to be associated with actively transcribed chromatin. By means of indirect immunofluorescence, it was shown to react with the nuclei of mouse fibroblasts and of brain cells from Xenopus and Drosophila, but not of Xenopus erythrocytes. The antibody was used to detect immunologically related proteins in giant chromosomes of the midge, Chironomus pallidivittatus. Indirect immunofluorescence with anti-HMG 14 antibody in polytene nuclei was restricted to the active puffs. Giant puffs (Balbiani rings) exhibited especially intense fluorescence in their peripheral regions. An inducible puff site, the Balbiani ring 6 locus, showed no reaction with the antibody prior to induction. When puff formation began, the chromosome site assumed a very intense fluorescence, which disappeared again when the Balbiani ring was recondensed. — Protein extracts of salivary gland nuclei were found on immunoblots to contain one major protein fraction that reacted with the anti-HMG 14 antibody. The electrophoretic mobility of this fraction was similar to that of calf thymus HMG 17. — It is concluded that actively transcribed puffs in polytene chromosomes contain HMG 14-related protein(s) that are not present in potentially active gene loci prior to induction.

Seventy-Five Years of Developmental Genetics: Ernst Caspari's Early Experiments on Insect Eye Pigmentation, Performed in an Academic Environment of Political Suppression

Anecdotal, Historical and Critical Commentaries on Genetics Edited by Adam Wilkins ERNST Caspari was the first researcher to use methods of developmental biology to analyze the action of a gene. By transplanting larval tissue between the wild type and a red-eyed mutant of the moth, Ephestia, he

A blotting procedure which preserves specific protein-DNA interactions

A quick, quantitative, and nonselective electrophoretic transfer of proteins from acetic acid-urea gels onto nitrocellulose, which preserves their ability to interact specifically with DNA, is achieved when exposure to dodecyl sulfate ions is avoided and a special type of nitrocellulose is used which contains cellulose phosphate ester. Filter-adsorbed histone H1 and other nuclear proteins from an insect, Chironomus thummi, were tested for binding of an AT-rich DNA sequence from the heterochromatin of the same organism under competitive conditions. On the blots, histone H1 exhibited the dependency of DNA binding on NaCl concentration and the preference for AT-rich DNA or poly[d(A-T)] found in quantitative filter-binding studies. By stepwise alteration of the NaCl molarity and competing Escherichia coli DNA concentration, respectively, in the binding buffer, two minor protein fractions could be identified in the heterogeneous extracts, one of which bound preferentially to AT-rich DNA, and the other bound to this sequence at up to 500 mM NaCl. Exposure to dodecyl sulfate led to a disappearance of the ability of these proteins to interact specifically with DNA. While nondenaturing transfer by diffusion (L. Levinger and A. Varshavsky (1982) Proc. Natl. Acad. Sci. USA 79, 7152) is a procedure that requires about 2 days, the present technique of gentle protein transfer for DNA binding studies requires only 2 to 3 h.

Transplantation of insect giant chromosome nuclei into amphibian oocytes.

Polytene salivary gland nuclei of Chironomus pallidivittatus were transplanted into oocytes of Xenopus laevis which were then cultured in vitro for 18 h. The giant chromosomes and nucleoli as well as the entire nuclei enlarged considerably in volume during this time. The polyteny and specific chromomere pattern of the chromosomes were maintained, and the puffing of the salivary gland-specific Balbiani rings was not noticeably changed. — Polytene nuclei from differentiated insect cells transplanted into Xenopus oocytes thus appear suited for exposing giant chromosomes in vivo to purified factors such as regulatory molecules.

In Memoriam Jan-Erik Edström (1931–2013)

JAN-ERIK Edstrom, an innovative pioneer in micromanipulation methods to study gene expression, died on August 30, 2013, in his home in Lomma, Sweden, after a long illness. He was born on April 21, 1931, in Gothenburg, Sweden. He studied medicine at the Medical School of Gothenburg’s Technical