Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes

doi:10.1016/S0092-8674(80)80048-1

Cell

Volume 19, Issue 3, March 1980, Pages 717-728

https://doi.org/10.1016/S0092-8674(80)80048-1 Get rights and content

Summary

A factor necessary for the accurate transcription of cloned Xenopus 5S genes in vitro has been isolated from soluble extracts of X. Iaevis ovaries. The activity of the factor was monitored by its ability to facilitate transcription of exogenous 5S genes in unfertilized egg extracts which are otherwise incompetent for 5S gene transcription. The factor was purified via ion exchange chromatography, and apparently consists of a 37,000 dalton polypeptide. This factor is necessary for the transcription of both the oocyte-type and somatic-type 5S genes of Xenopus, but is not required for, and has no detectable effect upon, the transcription of a cloned Xenopus tRNA₁^Met gene. The site of action of the factor has been investigated using the “footprinting” method of Galas and Schmitz (1978). The factor binds specifically to intragenic regions extending, approximately, from nucleotide positions 45 to 96 on both somatic and oocyte-type 5S genes. Additionally, this binding occurs independently of, and is not altered by, the presence of purified RNA polymerase III or unfertilized egg extracts. The probable role of this factor in transcription initiation is discussed.

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Distinct types of 5S rDNA repeats, differing by size and composition of its non-transcribed spacer (NTS), have been found in diverse taxa. Both concerted evolution and birth-and-death evolution have proven to play important roles in the evolution of the 5S rDNA family. In anurans, however, this subject has been underexplored as only a few anuran species had their 5S rDNA characterized and evolutionary analyzed to date. In the present study, we characterized the 5S rDNA sequences from species of two anuran families using classical molecular biology techniques and bioinformatic approaches. Based on the NTS, more than one type of 5S rDNA was identified in each analyzed species, which suggests that birth-and-death processes take part in the evolution of these sequences along the Anura tree of life. In addition, closely related species shared the same types of sequences, in accordance with the model of concerted evolution. We also found evidence of recombination between 5S rDNA and PcP190 satellite DNA, a repetitive sequence that is derived from the 5S rDNA. The interplay between 5S rDNA and PcP190 satellite DNA might favor the maintenance of the latter in the genome and respond to its presence in several species of frogs. The analysis of 5S rRNA transcripts confirmed the type I 5S rDNA of leiuperine as a functional 5S rRNA gene. Finally, the chromosome mapping of 5S rDNA sequences allowed some inferences of chromosome homology in Leiuperinae. In conclusion, our study provides additional information about the organization, differentiation and functionality of 5S rDNA in anuran species, revealing the potential participation of satellite DNA in the evolution of this family of rDNA.
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Citation Excerpt :
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Summary

References (29)

A control region in the center of the 5S RNA gene directs specific initiation of transcription: II. The 3′ border of the region

Cell

The in vitro transcription of 5S DNA

Separation and characterization of the subunits of ribonucleic acid polymerase

J. Biol. Chem.

Sequence organization of a cloned tDNA₁^met fragment from Xenopus laevis

Cell

Purification, properties and subunit structure of DNA-dependent RNA polymerase III from adenovirus 2-infected KB cells

J. Biol. Chem.

Nucleotide sequence of Xenopus borealis oocyte 5S DNA: comparison of sequences that flank several related eucaryotic genes

Cell

Assembly of SV40 chromatin in a cell-free system from Xenopus eggs

Cell

Selective transcription of the 5S genes in isolated chromatin by RNA polymerase III

Multiple forms of DNA-dependent RNA polymerase in Xenopus laevis: levels of activity during oocyte and embryonic development

J. Biol. Chem.

Factors involved in the transcription of purified genes by RNA polymerase III

Interaction of Int protein with specific sites on λ att DNA

Cell

A control region in the center of the 5S RNA gene directs specific initiation of transcription: I. The 5′ border of the region

Cell

The use of thin acrylamide gels for DNA sequencing

FEBS Letters

Purification and subunit structure of DNA-dependent RNA polymerase I from the mouse myeloma

J. Biol. Chem.

Cited by (467)

Transcription elongation mechanisms of RNA polymerases I, II, and III and their therapeutic implications

Structural basis of TFIIIC-dependent RNA polymerase III transcription initiation

High diversity of 5S ribosomal DNA and evidence of recombination with the satellite DNA PcP190 in frogs

Snapshots of RNA polymerase III in action – A mini review

Fe-S clusters masquerading as zinc finger proteins

Eukaryotic Transcription Turns 50