Abstract
Peri-centromeric regions of Drosophila melanogaster chromosomes appear heterochromatic in mitotic cells and become greatly underrepresented in giant polytene chromosomes, where they aggregate into a central mass called the chromocenter. We used P elements inserted at sites dispersed throughout much of the mitotic heterochromatin to analyze the fate of 31 individual sites during polytenization. Analysis of DNA sequences flanking many of these elements revealed that middle repetitive or unique sequence DNAs frequently are interspersed with satellite DNAs in mitotic heterochromatin. All nine Y chromosome sites tested were underrepresented >20-fold on Southern blots of polytene DNA and were rarely or never detected by in situ hybridization to salivary gland chromosomes. In contrast, nine tested insertions in autosomal centromeric heterochromatin were represented fully in salivary gland DNA, despite the fact that at least six were located proximal to known blocks of satellite DNA. The inserted sequences formed diverse, site-specific morphologies in the chromocenter of salivary gland chromosomes, suggesting that domains dispersed at multiple sites in the centromeric heterochromatin of mitotic chromosomes contribute to polytene β-heterochromatin. We suggest that regions containing heterochromatic genes are organized into dispersed chromatin configurations that are important for their function in vivo.
Full Text
The Full Text of this article is available as a PDF (9.5 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abad J. P., Carmena M., Baars S., Saunders R. D., Glover D. M., Ludeña P., Sentis C., Tyler-Smith C., Villasante A. Dodeca satellite: a conserved G+C-rich satellite from the centromeric heterochromatin of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4663–4667. doi: 10.1073/pnas.89.10.4663. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baiborodin S. I., Baricheva E. M., Bogachev S. S., Borisevich I. V., Strotz O. V., Filippova M. A., Sharakhov I. V., Shilov A. G. A molecular and cytogenetic analysis of lambda 20p7 fragment DNA from the proximal beta-heterochromatin of Drosophila melanogaster. Gene. 1993 Dec 8;134(2):175–181. doi: 10.1016/0378-1119(93)90091-g. [DOI] [PubMed] [Google Scholar]
- Balakireva M. D., Shevelyov YuYa, Nurminsky D. I., Livak K. J., Gvozdev V. A. Structural organization and diversification of Y-linked sequences comprising Su(Ste) genes in Drosophila melanogaster. Nucleic Acids Res. 1992 Jul 25;20(14):3731–3736. doi: 10.1093/nar/20.14.3731. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Biggs W. H., 3rd, Zavitz K. H., Dickson B., van der Straten A., Brunner D., Hafen E., Zipursky S. L. The Drosophila rolled locus encodes a MAP kinase required in the sevenless signal transduction pathway. EMBO J. 1994 Apr 1;13(7):1628–1635. doi: 10.1002/j.1460-2075.1994.tb06426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonaccorsi S., Lohe A. Fine mapping of satellite DNA sequences along the Y chromosome of Drosophila melanogaster: relationships between satellite sequences and fertility factors. Genetics. 1991 Sep;129(1):177–189. doi: 10.1093/genetics/129.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brunner D., Oellers N., Szabad J., Biggs W. H., 3rd, Zipursky S. L., Hafen E. A gain-of-function mutation in Drosophila MAP kinase activates multiple receptor tyrosine kinase signaling pathways. Cell. 1994 Mar 11;76(5):875–888. doi: 10.1016/0092-8674(94)90362-x. [DOI] [PubMed] [Google Scholar]
- Caizzi R., Caggese C., Pimpinelli S. Bari-1, a new transposon-like family in Drosophila melanogaster with a unique heterochromatic organization. Genetics. 1993 Feb;133(2):335–345. doi: 10.1093/genetics/133.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooley L., Kelley R., Spradling A. Insertional mutagenesis of the Drosophila genome with single P elements. Science. 1988 Mar 4;239(4844):1121–1128. doi: 10.1126/science.2830671. [DOI] [PubMed] [Google Scholar]
- Devlin R. H., Bingham B., Wakimoto B. T. The organization and expression of the light gene, a heterochromatic gene of Drosophila melanogaster. Genetics. 1990 May;125(1):129–140. doi: 10.1093/genetics/125.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devlin R. H., Holm D. G., Morin K. R., Honda B. M. Identifying a single-copy DNA sequence associated with the expression of a heterochromatic gene, the light locus of Drosophila melanogaster. Genome. 1990 Jun;33(3):405–415. doi: 10.1139/g90-062. [DOI] [PubMed] [Google Scholar]
- Dimitri P. Cytogenetic analysis of the second chromosome heterochromatin of Drosophila melanogaster. Genetics. 1991 Mar;127(3):553–564. doi: 10.1093/genetics/127.3.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eberl D. F., Duyf B. J., Hilliker A. J. The role of heterochromatin in the expression of a heterochromatic gene, the rolled locus of Drosophila melanogaster. Genetics. 1993 May;134(1):277–292. doi: 10.1093/genetics/134.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gall J. G., Cohen E. H., Polan M. L. Reptitive DNA sequences in drosophila. Chromosoma. 1971;33(3):319–344. doi: 10.1007/BF00284948. [DOI] [PubMed] [Google Scholar]
- Gatti M., Pimpinelli S. Functional elements in Drosophila melanogaster heterochromatin. Annu Rev Genet. 1992;26:239–275. doi: 10.1146/annurev.ge.26.120192.001323. [DOI] [PubMed] [Google Scholar]
- Glaser R. L., Karpen G. H., Spradling A. C. Replication forks are not found in a Drosophila minichromosome demonstrating a gradient of polytenization. Chromosoma. 1992 Dec;102(1):15–19. doi: 10.1007/BF00352285. [DOI] [PubMed] [Google Scholar]
- Hayashi S., Ruddell A., Sinclair D., Grigliatti T. Chromosomal structure is altered by mutations that suppress or enhance position effect variegation. Chromosoma. 1990 Oct;99(6):391–400. doi: 10.1007/BF01726690. [DOI] [PubMed] [Google Scholar]
- Hilliker A. J., Appels R., Schalet A. The genetic analysis of D. melanogaster heterochromatin. Cell. 1980 Oct;21(3):607–619. doi: 10.1016/0092-8674(80)90424-9. [DOI] [PubMed] [Google Scholar]
- Hilliker A. J. Genetic analysis of the centromeric heterochromatin of chromosome 2 of Drosophila melanogaster: deficiency mapping of EMS-induced lethal complementation groups. Genetics. 1976 Aug;83(4):765–782. doi: 10.1093/genetics/83.4.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karpen G. H., Spradling A. C. Analysis of subtelomeric heterochromatin in the Drosophila minichromosome Dp1187 by single P element insertional mutagenesis. Genetics. 1992 Nov;132(3):737–753. doi: 10.1093/genetics/132.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karpen G. H., Spradling A. C. Reduced DNA polytenization of a minichromosome region undergoing position-effect variegation in Drosophila. Cell. 1990 Oct 5;63(1):97–107. doi: 10.1016/0092-8674(90)90291-l. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lakhotia S. C., Jacob J. EM autoradiographic studies on polytene nuclei of Drosophila melanogaster. II. Organization and transcriptive activity of the chromocentre. Exp Cell Res. 1974 Jun;86(2):253–263. doi: 10.1016/0014-4827(74)90711-3. [DOI] [PubMed] [Google Scholar]
- Levis R. W., Ganesan R., Houtchens K., Tolar L. A., Sheen F. M. Transposons in place of telomeric repeats at a Drosophila telomere. Cell. 1993 Dec 17;75(6):1083–1093. doi: 10.1016/0092-8674(93)90318-k. [DOI] [PubMed] [Google Scholar]
- Lohe A. R., Brutlag D. L. Multiplicity of satellite DNA sequences in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1986 Feb;83(3):696–700. doi: 10.1073/pnas.83.3.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lohe A. R., Hilliker A. J., Roberts P. A. Mapping simple repeated DNA sequences in heterochromatin of Drosophila melanogaster. Genetics. 1993 Aug;134(4):1149–1174. doi: 10.1093/genetics/134.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miklos G. L., Cotsell J. N. Chromosome structure at interfaces between major chromatin types: alpha- and beta-heterochromatin. Bioessays. 1990 Jan;12(1):1–6. doi: 10.1002/bies.950120102. [DOI] [PubMed] [Google Scholar]
- Miklos G. L., Yamamoto M. T., Davies J., Pirrotta V. Microcloning reveals a high frequency of repetitive sequences characteristic of chromosome 4 and the beta-heterochromatin of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2051–2055. doi: 10.1073/pnas.85.7.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mitchelson A., Simonelig M., Williams C., O'Hare K. Homology with Saccharomyces cerevisiae RNA14 suggests that phenotypic suppression in Drosophila melanogaster by suppressor of forked occurs at the level of RNA stability. Genes Dev. 1993 Feb;7(2):241–249. doi: 10.1101/gad.7.2.241. [DOI] [PubMed] [Google Scholar]
- Montell D. J., Rorth P., Spradling A. C. slow border cells, a locus required for a developmentally regulated cell migration during oogenesis, encodes Drosophila C/EBP. Cell. 1992 Oct 2;71(1):51–62. doi: 10.1016/0092-8674(92)90265-e. [DOI] [PubMed] [Google Scholar]
- Parks S., Wieschaus E. The Drosophila gastrulation gene concertina encodes a G alpha-like protein. Cell. 1991 Jan 25;64(2):447–458. doi: 10.1016/0092-8674(91)90652-f. [DOI] [PubMed] [Google Scholar]
- Pavan C. Two Types of Heterochromatin in Drosophila Nebulosa. Proc Natl Acad Sci U S A. 1946 May;32(5):137–145. doi: 10.1073/pnas.32.5.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pimpinelli S., Dimitri P. Cytogenetic analysis of segregation distortion in Drosophila melanogaster: the cytological organization of the Responder (Rsp) locus. Genetics. 1989 Apr;121(4):765–772. doi: 10.1093/genetics/121.4.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin G. M., Spradling A. C. Vectors for P element-mediated gene transfer in Drosophila. Nucleic Acids Res. 1983 Sep 24;11(18):6341–6351. doi: 10.1093/nar/11.18.6341. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spradling A. C. Position effect variegation and genomic instability. Cold Spring Harb Symp Quant Biol. 1993;58:585–596. doi: 10.1101/sqb.1993.058.01.065. [DOI] [PubMed] [Google Scholar]
- Spradling A., Orr-Weaver T. Regulation of DNA replication during Drosophila development. Annu Rev Genet. 1987;21:373–403. doi: 10.1146/annurev.ge.21.120187.002105. [DOI] [PubMed] [Google Scholar]
- Steinemann M., Steinemann S. Degenerating Y chromosome of Drosophila miranda: a trap for retrotransposons. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7591–7595. doi: 10.1073/pnas.89.16.7591. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steller H., Pirrotta V. P transposons controlled by the heat shock promoter. Mol Cell Biol. 1986 May;6(5):1640–1649. doi: 10.1128/mcb.6.5.1640. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Traverse K. L., Pardue M. L. Studies of He-T DNA sequences in the pericentric regions of Drosophila chromosomes. Chromosoma. 1989 Jan;97(4):261–271. doi: 10.1007/BF00371965. [DOI] [PubMed] [Google Scholar]
- Yamamoto M. T., Mitchelson A., Tudor M., O'Hare K., Davies J. A., Miklos G. L. Molecular and cytogenetic analysis of the heterochromatin-euchromatin junction region of the Drosophila melanogaster X chromosome using cloned DNA sequences. Genetics. 1990 Aug;125(4):821–832. doi: 10.1093/genetics/125.4.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young B. S., Pession A., Traverse K. L., French C., Pardue M. L. Telomere regions in Drosophila share complex DNA sequences with pericentric heterochromatin. Cell. 1983 Aug;34(1):85–94. doi: 10.1016/0092-8674(83)90138-1. [DOI] [PubMed] [Google Scholar]
- Young M. W. Middle repetitive DNA: a fluid component of the Drosophila genome. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6274–6278. doi: 10.1073/pnas.76.12.6274. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang P., Spradling A. C. Insertional mutagenesis of Drosophila heterochromatin with single P elements. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3539–3543. doi: 10.1073/pnas.91.9.3539. [DOI] [PMC free article] [PubMed] [Google Scholar]
