CHEMICAL SECONDARY STRUCTURE PROBING OF 2 HIGHLY METHYLATED REGIONS IN XENOPUS-LAEVIS 28S RIBOSOMAL-RNA

被引:4
作者
AJUH, PM [1 ]
MADEN, EB [1 ]
机构
[1] UNIV LIVERPOOL,DEPT BIOCHEM,LIVERPOOL L69 3BX,ENGLAND
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 1994年 / 1219卷 / 01期
关键词
LARGE RIBOSOMAL SUBUNIT RNA; 2'-O-METHYLATION; SECONDARY STRUCTURE; RIBOSOMAL RNA; RNA STRUCTURE; METHYLATION; (X-LAEVIS);
D O I
10.1016/0167-4781(94)90250-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The large ribosomal subunit (LSU) RNA or 28S rRNA of vertebrates is characterized by two highly conserved and methylated regions towards the 3' end of the molecule that extend from domains IV to V of the molecule. In this report we describe the probing of the secondary structure of these two highly methylated regions in Xenopus laevis LSU RNA by chemical modification using the single-strand nucleotide specific probes; dimethyl sulphate (DMS) and 1-cyclo-hexyl-3-(2-morpholinoethyl-carbodiimide metho-p-toluene sulphonate (CMCT) followed by primer extension. The same regions in in vitro synthesized unmethylated X. laevis 28S rRNA were also probed for comparison. Our results in general tend to support the theoretically determined secondary structure model for the probed domains. From the results obtained, methylated cellular LSU RNA appears to be relatively more reactive than the in vitro transcript to the chemical probes. Accessibility to the probes was found to be similar at most sites for cellular and in vitro transcript LSU RNAs. This implies that structural destabilization due to 2'-O-methylations in cellular LSU RNA is not significant.
引用
收藏
页码:89 / 97
页数:9
相关论文
共 24 条
[1]   XENOPUS-BOREALIS AND XENOPUS-LAEVIS 28S RIBOSOMAL DNA AND THE COMPLETE 40S RIBOSOMAL PRECURSOR RNA CODING UNITS OF BOTH SPECIES [J].
AJUH, PM ;
HEENEY, PA ;
MADEN, BEH .
PROCEEDINGS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 1991, 245 (1312) :65-71
[2]  
AJUH PM, 1991, THESIS U LIVERPOOL L
[3]  
AKHTAR Y, 1987, THESIS U LIVERPOOL L
[4]  
CARBOCHE M, 1977, EUR J BIOCHEM, V74, P19
[5]   ISOLATION OF BIOLOGICALLY-ACTIVE RIBONUCLEIC-ACID FROM SOURCES ENRICHED IN RIBONUCLEASE [J].
CHIRGWIN, JM ;
PRZYBYLA, AE ;
MACDONALD, RJ ;
RUTTER, WJ .
BIOCHEMISTRY, 1979, 18 (24) :5294-5299
[6]  
EGEBJERG J, 1990, RIBOSOME, P168
[7]   PROBING THE STRUCTURE OF RNAS IN SOLUTION [J].
EHRESMANN, C ;
BAUDIN, F ;
MOUGEL, M ;
ROMBY, P ;
EBEL, JP ;
EHRESMANN, B .
NUCLEIC ACIDS RESEARCH, 1987, 15 (22) :9109-9128
[8]   THE SECONDARY STRUCTURE OF HUMAN 28S RIBOSOMAL-RNA - THE STRUCTURE AND EVOLUTION OF A MOSAIC RIBOSOMAL-RNA GENE [J].
GORSKI, JL ;
GONZALEZ, IL ;
SCHMICKEL, RD .
JOURNAL OF MOLECULAR EVOLUTION, 1987, 24 (03) :236-251
[9]   A COMPILATION OF LARGE SUBUNIT RNA SEQUENCES PRESENTED IN A STRUCTURAL FORMAT [J].
GUTELL, RR ;
FOX, GE .
NUCLEIC ACIDS RESEARCH, 1988, 16 :R175-R269
[10]   CONSERVATION OF PRIMARY STRUCTURE AT 3' END OF 18S R-RNA FROM EUKARYOTIC CELLS [J].
HAGENBUCHLE, O ;
SANTER, M ;
STEITZ, JA ;
MANS, RJ .
CELL, 1978, 13 (03) :551-563