We demonstrate that in Drosophila melanogaster the histone H3.3 replacement variant is encoded by two genes, H3.3A and H3.3B. We have isolated cDNA clones for H3.3A and cDNA and genomic clones for H3.3B. The genes encode exactly the same protein but are widely divergent in their untranslated regions (UTR). Both genes are expressed in embryos and adults; they are expressed in the gonads as well as in somatic tissues of the flies. However, only one of them, H3.3A, shows strong testes expression. The 3' UTR of the H3.3A gene is relatively short (approximately 250 nucleotides (nt)). H3.3B transcripts can be processed at several polyadenylation sites, the longest with a 3' UTR of more than 1500 nt. The 3' processing sites, preferentially used in the gonads and somatic tissues, are different. We have also isolated the Drosophila hydei homologues of the two H3.3 genes. They are quite similar to the D. melanogaster genes in their expression patterns. However, in contrast to their vertebrate counterparts, which are highly conserved in their noncoding regions, the Drosophila genes display only limited sequence similarity in these regions.

The effects of the plant growth retardant tetcyclacis on in vitro tuber formation in potatoes was studied, using two different approaches : 1 . tuber formation in various lines that did not or hardly form tubers under control conditions, and 2 . tuber formation by the variety Bintje, which readily forms tubers . The ABA-deficient (droopy) lines of S. phureja hardly formed tubers without the addition of tetcyclacis . In the presence of this growth retardant tuberization was nearly 100%, within three weeks of in vitro culture, even in the absence of cytokinin . A series of somatic hybrids between S . tuberosum and S. brevidens, that did not form tubers in field and pot experiments, were tested . They all formed tubers in vitro in the presence of tetcyclacis . Stoloniferous shoots formed on single-node cuttings from in vitro grown Solanum tuberosum var Bintje plantlets were transferred to media containing a high level of sucrose . In the presence of tetcyclacis, tuber formation started after 4 days, reaching a maximum level of 80% at day 7 . Tubers formed in the presence of tetcyclacis, accumulated starch and expressed several tuber-specific genes . These effects were fully antagonized by gibberellic acid . It is concluded that the growth retardant tetcyclacis is a potent tool in the study of tuber formation in potatoes .Abbreviations : ABA = abscisic acid; BAP = benzylaminopurine ; GA3 = gibberellic acid ; STS = silver thiosulphate ; TET = tetcyclacis . 257

Searching for structural proteins involved in spermatogenesis of Drosophila, we found a novel myosin isoform in the testis of Drosophila hydei and D. melanogaster. The transcript encoding this isoform, which we called 'minor-myosin', initiates within the intron between exons 12 and 13 of the muscle myosin heavy chain (mMHC) gene. Minor-myosin contains a common myosin tail but no ordinary myosin head domain. Instead, it has a short N-terminal domain which displays similarity with the N-termini of certain myosin light chain proteins. Western blots with male germ line mutants showed that the novel mMHC isoform is synthesized in the male germ cells, mainly postmeiotically. However, minor-myosin is not testis-specific, as it is expressed at a low level in the fly carcasses. The possible functions of the myosin isoform in the male germ line are discussed.

The muscle-myosin heavy-chain (mMHC) gene of Drosophila hydei has been sequenced completely (size 23.3 kb). The sequence comparison with the D. melanogaster mMHC gene revealed that the exon-intron pattern is identical. The protein coding regions show a high degree of conservation (97%). The alternatively spliced exons (3a-b, 7a-d, 9a-c, 11a-e, and 15a-b) display more variations in the number of nonsynonymous and synonymous substitutions than the common exons (2, 4, 5, 6, 8, 10, 12, 13, 14, 16, 17, and 19). The base composition at synonymous sites of fourfold degenerate codons (third position) is not biased in the alternative exons. In the common exons there exists a bias for C and against A. These findings imply that the alternative exons of the Drosophila mMHC gene evolve at a different, in several cases higher, rate than the common ones. The 5' splice junctions and 5' and 3' untranslated regions show a high level of similarity, indicating a functional constraint on these sequences. The intron regions vary considerably in length within one species, but the corresponding introns are very similar in length between the two species and all contain stretches of sequence similarity. A particular example is the first intron, which contains multiple regions of similarity. In the conserved regions of intron 12 (head-tail border) sequences were found which have the potential to direct another smaller mMHC transcript.

We have found that defective gypsy retrotransposons are a major constituent of the lampbrush loop pair Nooses in the short arm of the Y chromosome of Drosophila hydei. The loop pair is formed by male fertility gene Q during the primary spermatocyte stage of spermatogenesis, each loop being a single transcription unit with an estimated length of 260 kb. Using fluorescent in situ hybridization, we show that throughout the loop transcripts gypsy elements are interspersed with blocks of a tandemly repetitive Y-specific DNA sequence, ay1. Nooses transcripts containing both sequence types show a wide size range on Northern blots, do not migrate to the cytoplasm, and are degraded just before the first meiotic division. Only one strand of ay1 and only the coding strand of gypsy can be detected in the loop transcripts. However, as cloned genomic DNA fragments also display opposite orientations of ay1 and gypsy, such DNA sections cannot be part of the Nooses. Hence, they are most likely derived from the flanking heterochromatin. The direction of transcription of ayl and gypsy thus appears to be of a functional significance.