Studies on Nucleolar RNA Synthesis in Drosophila Melanogaster

1972 ◽  
Vol 11 (3) ◽  
pp. 689-697
Author(s):  
H. M. KRIDER ◽  
W. PLAUT

The influence of conditions resulting in bobbed phenotypes on nucleolar RNA synthesis and the formation of constrictions at nucleolus organizers was examined in larval tissues of Drosophila melanogaster. By means of [3H]uridine incorporation and autoradiographic analysis, a mutation at the bobbed locus was shown to limit the rate of nucleolar RNA synthesis in salivary glands of XO larvae. The formation of constrictions at the organizer sites of a 4-nucleolus-organizer stock was monitored in dividing neuroblast cells stained with acridine orange. Loss of the ribosomal cistrons had been reported by other workers when such stocks were maintained for several generations. In the first generation in our work, constrictions were visible at only 2 of the 4 nucleolus organizers. This situation persisted until the fifth generation, when constrictions appeared at all 4 of the organizer sites. An increase in the rate of nucleolar RNA synthesis in the salivary glands was temporally correlated with the appearance of the extra constrictions. We interpret these observations to mean that 2 of the organizers of the 4-nucleolus-organizer stock were caused to function through the loss of ribosomal RNA cistrons; thus the functional status of an organizer would appear to be subject to control.

1972 ◽  
Vol 11 (3) ◽  
pp. 675-687
Author(s):  
H. M. KRIDER ◽  
W. PLAUT

Nucleolar RNA synthesis in salivary glands from XX, XY, and XO larvae having different numbers of nucleolus organizers was examined autoradiographically following incubation of the tissues in [3H]uridine. In addition, the presence or absence of secondary constrictions was monitored in neuroblast preparations using acridine orange staining. It was observed that: (1) The rate of nucleolar [3H]uridine incorporation is independent of the number of nucleolus organizers in the cell; (2) nucleolar incorporation in XO tissues is elevated relative to that observed in XX or XY material; and (3) where the number of organizers is in excess of the wild type (2), secondary constrictions form at only 2 of the nucleolus organizer sites. From these and related observations we suggest that there are 2 forms of transcriptional control for nucleolar RNA synthesis. One acts as an on-off control, influencing the formation of secondary constrictions at the nucleolus organizers. The other form modulates rates of transcription at organizers where constrictions have been formed.


1969 ◽  
Vol 14 (2) ◽  
pp. 137-150 ◽  
Author(s):  
S. C. Lakhotia ◽  
A. S. Mukherjee

Morphology and the rate of RNA synthesis of the X-chromosome in XX/XO mosaic larval salivary glands of Drosophila melanogaster have been examined. For this purpose the unstable ring-X was utilized to produce XX and XO nuclei in the same pair of glands. The width of the X-chromosome and the left arm of the 3rd chromosome (3L) of larval salivary glands was measured and the rate of RNA synthesis by them was studied upon the use of [3H]uridine autoradiography in such XX (female) and XO (male) nuclei developing in a female background (i.e. otherwise genotypically XX). In such mosaic glands the width of the single X-chromosome of male nuclei is nearly as great as that of the paired two X's of female nuclei, as is also the case in normal male (X Y) and female (XX). The single X of male nuclei synthesizes RNA at a rate equal to that of the paired two X's of female nuclei and nearly twice that of an unpaired X of XX nuclei. Neither the developmental physiology of the sex nor the proportion of XO nuclei in a pair of mosaic salivary glands of an XX larva has any influence on these two characteristics of the male X-chromosome.It is suggested that dosage compensation in Drosophila is achieved chiefly, if not fully, by a hyperactivity of the male X, in contrast to the single X inactivation in female mammals, that this hyperactivity of the male X is expressed visibly in the morphology and metabolic activity of the X-chromosome in the larval salivary glands of the male, and that this hyperactivity and therefore dosage compensation in Drosophila in general is not dependent on sex-differentiation, but is a function of the doses of the X-chromosome itself.


Genetics ◽  
1987 ◽  
Vol 116 (2) ◽  
pp. 241-251
Author(s):  
M R Gillings ◽  
R Frankham ◽  
J Speirs ◽  
M Whalley

ABSTRACT The nucleolus organizers on the X and Y chromosomes of Drosophila melanogaster are the sites of 200-250 tandemly repeated genes for ribosomal RNA. As there is no meiotic crossing over in male Drosophila, the X and Y chromosomal rDNA arrays should be evolutionarily independent, and therefore divergent. The rRNAs produced by X and Y are, however, very similar, if not identical. Molecular, genetic and cytological analyses of a series of X chromosome rDNA deletions (bb alleles) showed that they arose by unequal exchange through the nucleolus organizers of the X and Y chromosomes. Three separate exchange events generated compound X·YL chromosomes carrying mainly Y-specific rDNA. This led to the hypothesis that X-Y exchange is responsible for the coevolution of X and Y chromosomal rDNA. We have tested and confirmed several of the predictions of this hypothesis: First, X·YL chromosomes must be found in wild populations. We have found such a chromosome. Second, the X·YL chromosome must lose the YL arm, and/or be at a selective disadvantage to normal X+ chromosomes, to retain the normal morphology of the X chromosome. Six of seventeen sublines founded from homozygous X·YLbb stocks have become fixed for chromosomes with spontaneous loss of part or all of the appended YL. Third, rDNA variants on the X chromosome are expected to be clustered within the X+ nucleolus organizer, recently donated ("Y") forms being proximal, and X-specific forms distal. We present evidence for clustering of rRNA genes containing Type 1 insertions. Consequently, X-Y exchange is probably responsible for the coevolution of X and Y rDNA arrays.


1977 ◽  
Vol 25 (1) ◽  
pp. 111-123
Author(s):  
B.T. Luck ◽  
E.G. Jordan

Stages of meiosis from the bluebell Endymion non-scriptus (L.) were studied by electron microscopy. The segregated components of the nucleolus at meiotic prophase underwent fragmentation and dissolution at pachytene-diplotene. Nucleoli were absent during both meiotic divisions and reformed on the nucleolus organizer into a fibrillar mass from scattered fibrillar components at the dyad and tetrad stages. Ti is argued that the fibrillar region shows continuity through nuclear division though undergoing structural transformations in the process. Nucleolar reformation occurs on condensed nucleolus organizers. Processing of the ribosomal precursors and the resumption of RNA synthesis is discussed in relation to the dispersal of the nucleolus organizer into the fibrillar region of the reformed nucleolus.


Genetics ◽  
1973 ◽  
Vol 73 (3) ◽  
pp. 429-434
Author(s):  
J James Donady ◽  
R L Seecof ◽  
M A Fox

ABSTRACT Drosophila melanogaster embryos that lacked ribosomal DNA were obtained from appropriate crosses. Cells were taken from such embryos before overt differentiation took place and were cultured in vitro. These cells differentiated into neurons and myocytes with the same success as did wild-type controls. Therefore, ribosomal RNA synthesis is not necessary for the differentiation of neurons and myocytes in vitro.


1965 ◽  
Vol 27 (3) ◽  
pp. 545-550 ◽  
Author(s):  
A. Di Marco ◽  
R. Silvestrini ◽  
S. Di Marco ◽  
T. Dasdia

The effect has been studied of Actinomycin D, Daunomycin (Da.), and Da. N acetyl derivative on mitotic activity and on the nucleic acid synthesis of in vitro HeLa cell cultures. The experiments were carried out by means of the radioautographic technique using stripping films. The relative uptake of thymidine-H3 and uridine-H3 was determined by means of the reduced silver grain count present in the nuclei of controls and treated cells. The mitotic activity and thymidine incorporation were noticeably reduced by Daunomycin and Actinomycin, whereas both processes appeared less affected by Da. N acetyl derivative. As regards nuclear RNA synthesis, all three antibiotics at low doses chiefly inhibit nucleolar RNA synthesis. On the other hand, whilst Actinomycin at higher doses causes an almost total inhibition of the synthesis of the whole nuclear RNA, in Daunomycin- and Da. N acetyl derivative-treated cells extranucleolar RNA synthesis is less susceptible to inhibition.


Genetics ◽  
1988 ◽  
Vol 120 (4) ◽  
pp. 1053-1059
Author(s):  
L G Robbins ◽  
E E Swanson

Abstract Rex-induced mitotic recombination was used to determine whether nucleolus organizers can pair in both inverted and noninverted orientations. Two target chromosomes, each duplicated for the rDNA region, were exposed to maternal Rex activity. Recombination in one orientation should yield deletion of the material between the two nucleolus organizers, recombination in the other orientation should yield inversion of the same material. Both products were recovered from both target chromosomes. The generality of using Rex-mediated recombination for analysis of the rDNA is considered.


1973 ◽  
Vol 133 (4) ◽  
pp. 797-804 ◽  
Author(s):  
Donner F. Babcock ◽  
Marvin A. Rich

1. In the spleens of infected mice, the Friend leukaemia virus induces a sharp increase in the ability of subsequently isolated nuclei to incorporate exogenous UTP into an acid-insoluble product. Inhibitor studies indicate that the incremental RNA synthesis proceeds from a DNA template and that both nucleolar and nucleoplasmic activities are involved. 2. The partially purified DNA-dependent RNA polymerases from control and virus-infected tissue are indistinguishable with respect to chromatographic mobility, dependence on bivalent cations, ionic strength, pH and their susceptibility to α-amanitin. The RNA polymerases of the murine spleen resemble the enzymes of other mammalian tissue in these properties. 3. A comparison of the amount of polymerase solubilized from normal and infected tissue correlates with the activity observed in assays of the respective nuclei. These experiments indicated that the increase in nucleolar RNA synthesis after infection is mediated by increased extractable polymerase I activity whereas the change in nucleoplasmic RNA synthesis results from an alteration of chromatin or a chromatin-associated factor.


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