The structural organization of ribosomal DNA in drosophila melanogaster

The genes coding for the large rRNAs (rDNA) were purified from Drosophila melanogaster embryos by two different methods. Purified rDNA was analyzed by gel electrophoresis after digestion with restriction endonucleases and by electron microscopy. Each repeating unit of rDNA consists of a gene region that codes for a transcript which includes the 18S and 28S rRNA sequences plus a nontrascribed spacer. Analysis of rRNA/rDNA hybrids in the electron microscope reveals that two types of repeating units exist in the rDNA from this fly. One of these is characterized by insertions of various lengths within the 28S gene. Insertions range in size from about 0.5 kilo base pairs (kb) to 6.0 kb and occur in distinct size classes which are multiples of 0.5 kb. Figure 10 summarizes the structure of the rDNA repeating unit. Repeats with insertions constitute about two thirds, and repeats without insertions about one third of all repeating units in the sample studied where 86% of the rDNA is derived from the X chromosome nucleolus organizer. Statistical analysis of pairs of of nearest-neighbor repeats demonstrates that repeats with and without insertions are interspersed in the X chromosome nucleolus organizer, and that scrambling is close to that expected for random arrangement. Insertions are the major source of length heterogeneity in this rDNA. A minor length heterogeneity also exists in thie nontranscribed spacer of Drosophila rDNA as demonstrated by heteroduplex mapping of restriction fragments in the electron microscope.

[1]  G. P. Smith,et al.  Unequal crossover and the evolution of multigene families. , 1974, Cold Spring Harbor symposia on quantitative biology.

[2]  David M. Glover,et al.  A novel arrangement of the 18S and 28S sequences in a repeating unit of drosophila melanogaster rDNA , 1977, Cell.

[3]  E. Southern Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[4]  B. Hamkalo,et al.  Ultrastructure of active eukaryotic genomes. , 1974, Cold Spring Harbor symposia on quantitative biology.

[5]  U. Loening The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophresis. The effects of changes in conformation. , 1969, The Biochemical journal.

[6]  R. Perry,et al.  The 5 s RNA genes of Drosophila melanogaster. , 1970, Journal of molecular biology.

[7]  Jerry Manning,et al.  Sequence arrangement of the rDNA of Drosophila melanogaster , 1977, Cell.

[8]  K. Tartof Increasing the Multiplicity of Ribosomal RNA Genes in Drosophila melanogaster , 1971, Science.

[9]  I. Dawid,et al.  The molecular basis for length heterogeneity in ribosomal DNA from Xenopus laevis. , 1976, Journal of molecular biology.

[10]  M. Wu,et al.  Expression of the mitochondrial genome in HeLa cells. XIV. The relative positions of the 4 S RNA genes and of the ribosomal RNA genes in mitochondrial DNA. , 1972, Journal of molecular biology.

[11]  I. Dawid,et al.  Similarities and differences in the structure of X and Y chromosome rRNA genes of Drosophila , 1976, Nature.

[12]  J. Greenberg Synthesis and properties of ribosomal RNA in Drosophila. , 1969, Journal of molecular biology.

[13]  M. Fischberg,et al.  Localization of the ribosomal DNA complements in the nucleolar organizer region of Xenopus laevis. , 1966, National Cancer Institute monograph.

[14]  S. L. Commerford Iodination of nucleic acids in vitro. , 1971, Biochemistry.

[15]  B. Maden,et al.  Nature of the ribosomal RNA transcribed from the X and Y chromosomes of Drosophila melanogaster. , 1974, Journal of molecular biology.

[16]  S. McKnight,et al.  Ultrastructural patterns of RNA synthesis during early embryogenesis of Drosophila melanogaster , 1976, Cell.

[17]  N. Davidson,et al.  Separation of Deoxyribonucleic Acids by Hg(II) Binding and Cs2SO4 Density-Gradient Centrifugation* , 1965 .

[18]  D. D. Brown,et al.  Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4 s RNA, 5 s RNA and ribosomal RNA. , 1968, Journal of molecular biology.

[19]  I. Dawid,et al.  Arrangement of length heterogeneity in repeating units of amplified and chromosomal ribosomal DNA from Xenopus laevis. , 1976, Journal of molecular biology.

[20]  W. Prensky,et al.  The use of iodinated RNA for gene localization. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[21]  I. Dawid Mitochondrial RNA In Xenopus laevis. I. The expression of the mitochondrial genome. , 1972, Journal of molecular biology.

[22]  I. Dawid,et al.  Patterns of ribosomal DNA spacer lengths are inherited. , 1976, Journal of molecular biology.

[23]  S. Spiegelman,et al.  On the chromosomal distribution of DNA complementary to ribosomal and soluble RNA. , 1966, National Cancer Institute monograph.

[24]  H. Goodman,et al.  Specificity of substrate recognition by the EcoRI restriction endonuclease. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. Deutch,et al.  Amplified ribosomal DNA from Xenopus laevis has heterogeneous spacer lengths. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Raymond L. White,et al.  Characterization of six cloned DNAs from drosophila melanogaster, including one that contains the genes for rRNA , 1975, Cell.

[27]  F. Ritossa Crossing-over between X AND Y chromosomes during ribosomal DNA magnification in Drosophila melanogaster. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[28]  B. Jacq,et al.  Late steps in the maturation of Drosophila 26 S ribosomal RNA: generation of 5-8 S and 2 S RNAs by cleavages occurring in the cytoplasm. , 1976, Journal of molecular biology.

[29]  Raymond L. White,et al.  R loop mapping of the 18S and 28S sequences in the long and short repeating units of drosophila melanogaster rDNA , 1977, Cell.

[30]  I. Dawid,et al.  Secondary structure maps of ribosomal RNA and DNA. I. Processing of Xenopus laevis ribosomal RNA and structure of single-stranded ribosomal DNA. , 1974, Journal of molecular biology.