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PMID: 806350 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Polypyrimidine segments in Drosophila melanogaster DNA: II. Chromosome location and nucleotide sequence.

Cell ·Vol. 5 ·No. 2 ·1975-06-00 ·Pages 183-94

Sederoff R, Lowenstein L

Abstract

Long pyrimidine tracts, purified from Drosophila melanogaster DNA after treatment with formic acid-diphenylamine, were used as template for E. coli RNA polymerase to produce a polynucleotide containing only purines. This polypurine RNA hybridized specifically to D. melanogaster DNA with high efficiency at low Cot values. The resulting hybrid showed high thermal stability. When polypurine RNA was subjected to complete hydrolysis with ribonuclease T1, over 90% of the nucleotide products were ApGp and ApApGp. Partial hydrolysis yielded a distinct additional component, ApApGpApGp + ApGpApApGp. We conclude that the major sequence in the polypurine transcript is (ApGpApApGp)n. In situ hybridization to salivary gland polytene chromosomes and to metaphase chromosomes from neural ganglia indicated that polypyrimidines complementary to polypurine RNA are located in heterochromatin. In femal cells, the predominant labeling was on centromeric heterochromatin of the 2nd chromosome. We have verified the location of polypyrimidines in neural ganglion cells, by using a cytological marker of chromosomes 2. In male cells, hybrid was also found on the Y chromosome.

MeSH Terms
Adenosine Triphosphate/metabolism Base Sequence Cell Line Cell-Free System Chromosomes/analysis DNA/analysis DNA-Directed RNA Polymerases/metabolism Drosophila melanogaster/analysis Escherichia coli/enzymology Ganglia/analysis Guanosine Triphosphate/metabolism Heterochromatin/analysis Nucleic Acid Hybridization Purine Nucleotides/biosynthesis Pyrimidine Nucleotides/analysis RNA/analysis,biosynthesis Salivary Glands/analysis Templates, Genetic Tritium
Chemicals
Heterochromatin Purine Nucleotides Pyrimidine Nucleotides Tritium RNA Guanosine Triphosphate Adenosine Triphosphate DNA DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sederoff R
Lowenstein L
Article Info
Journal
Cell
Abbr.
Cell
ISSN
0092-8674
Published
1975-06-00
Pages
183-94
Language
English
Region
United States
NLM ID
0413066
Subset
IM
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