Direct chemical probing of the conformation of the 3' functional domain of rabbit 18S rRNA in 40S subunits, 80S ribosomes and polyribosomes
Recent evidence suggests that the 3' minor domain of eukaryotic 18S rRNA, as in prokaryotes, is directly involved in protein biosynthesis. To determine regions of possible functional importance, they have probed the higher order structure of rabbit 18S rRNA in both 40S subunits and 80S ribosomes, as well as polyribosomes using the single-strand specific chemical probes dimethyl sulfate (DMS) and diethyl pyrocarbonate (DEPC) which react with unpaired guanosine and adenosine residues, respectively. The modified 18S rRNA was isolated from these particles and the resultant modified nucleotides identified on polyacrylamide sequencing gels upon either aniline-induced strand scission of /sup 32/P-end-labeled intact rRNA or by DNA primer extension using sequence specific deoxyoligomers with reverse transcriptase. Their results indicate a decreased reactivity of residue C-1692 in rabbit 18S rRNA (corresponding to C-1400 E. coli) within the putative tRNA contact site in polyribosomes as compared with 40S subunits and 80S ribosomes. They have also determined varying reactivities of a number of other residues within specific regions of the 3' functional domain when 40S, 80S, and polyribosomes are compared, which may be important for both subunit association as well as mRNA binding.
- Research Organization:
- Cornell Univ. Medical College, New York, NY
- OSTI ID:
- 6023443
- Report Number(s):
- CONF-870644-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PROTEINS
BIOSYNTHESIS
RIBOSOMAL RNA
MOLECULAR STRUCTURE
ADENOSINE
CARBONATES
ELECTROPHORESIS
GUANOSINE
PHOSPHORUS 32
RABBITS
RIBOSOMES
SULFATES
TRACER TECHNIQUES
ANIMALS
AROMATICS
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON COMPOUNDS
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
HETEROCYCLIC COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
NUCLEI
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANOIDS
OXYGEN COMPOUNDS
PHOSPHORUS ISOTOPES
PURINES
RADIOISOTOPES
RIBOSIDES
RNA
SULFUR COMPOUNDS
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques