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PMID: 6250536 Published · ppublish English Comparative Study Journal Article

Distribution of sequences common to the 25--28S-ribonucleic acid genes of Xenopus laevis and Neurospora crassa.

The Biochemical journal ·Vol. 187 ·No. 1 ·1980-04-01 ·Pages 75-90

Cox RA, Thompson RD

Abstract

The extent of homology between the nucleotide sequence of L-rRNA (the major RNA component of the larger ribosomal subparticle) of a lower eukaryote (Neurospora crassa) and an amphibian (Xenopus laevis) was investigated by utilizing rDNA (DNA coding for rRNA) of X. laevis cloned in plasmids pMB9 and pML2, and rDNA of N. crassa cloned in bacteriophage lambda. Hybridization studies revealed that sequences common to both N. crassa and X. laevis L-rRNA comprise a total of approx. 1050 /+- 200 nucleotides. The thermal stability of the X. laevis rDNA.N. crassa L-rRNA hybrid was 5 degrees C lower than that of the X. laevis rDNA.X. laevis L-rRNA duplex, indicating the presence of fewer than 10% mismatches in homologous sequences. X. laevis rDNA was analysed by means of restriction endonucleases and hybridization with 125I-labelled N. crassa L-rRNA. Most (at least 95%) of the conserved sequences were present in a 3000-base-pair fragment produced by restriction with endonucleases HindIII and BamHI. This fragment, which includes the 3'-OH terminus of the L-rRNA-coding region, was used as an adaptor in the construction of a bacteriophage-lambda recombinant. One section of the recombinant phage terminating in a HindIII-specific site was obtained from bacteriophage lambda plac5 (after restriction with endonuclease HindIII). A second section terminating in a BamHi-specific site was obtained from bacteriophage lambda 540 (after restriction with endonuclease BamHI). These two parts were joined by means of the X. laevis rDNA fragment. Further analysis of cloned rDNA by means of restriction endonucleases confirmed that conserved sequences were widely distributed throughout the 3000-base-pair fragment produced by HindIII and BamHi endonucleases. A 3400-base-pair fragment of N. crassa rDNA cloned in a bacteriophage lambda [Cox & Peden (1979) Mol. Gen. Genet. 174, 17--24] was restricted with endonucleases. The products were hybridized with 125I-labelled X. laevis L-rRNA. Conserved sequences were shown to be distributed over a range of approx. 1600--2700 base-pairs. Hence, in neither X. laevis nor N. crassa L-rRNA can be conserved sequences from a single block; instead regions of high and low (or no) homology must be intermingled. Both N. crassa rDNA and X. laevis rDNA were found to hybridize with Drosophila melanogaster L-rDNA sequences. Those rDNA fragments with sequences common to X. laevis and N. crassa L-rRNA also hybridized with D. melanogaster L-rRNA probe. Thus the same set of conserved sequences may be present in all three species.

MeSH Terms
Animals Base Sequence DNA DNA Restriction Enzymes Drosophila melanogaster Electrophoresis, Agar Gel Genes Neurospora/genetics Neurospora crassa/genetics Nucleic Acid Hybridization RNA, Ribosomal/genetics Species Specificity Xenopus/genetics
Chemicals
RNA, Ribosomal DNA DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cox R A
Thompson R D
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34 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1870075
Published
1980-04-01
Pages
75-90
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1162495
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1870075 PMC Full Text View journal details
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