Identification and assignment of base pairs in four helical segments of Bacillus megaterium ribosomal 5S RNA and its ribonuclease T1 cleavage fragments by means of 500-MHz proton homonuclear Overhauser enhancements

Journal Article · Tue Jan 23 04:00:00 EST 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00455a006· OSTI ID:6717768

Kim, J H; Marshall, A G ^[1]

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1. Ohio State Univ., Columbus (USA)

Three different fragments of Bacillus megaterium ribosomal 5S RNA have been produced by enzymatic cleavage with ribonuclease T1. Fragment A consists of helices II and III, fragment B contains helix IV, and fragment C contains helix I of the universal 5S rRNA secondary structure. All (eight) imino proton resonances in the downfield region (9-15 ppm) of the 500-MHz proton FT NMR spectrum of fragment B have been identified and assigned as G{sub 80}{center dot}C{sub 92}{center dot}G{sub 81}{center dot}C{sub 91}-G{sub 82}{center dot}C{sub 90}-A{sub 83}{center dot}U{sub 89}-C{sub 84}{center dot}G{sub 88} and three unpaired U's in helix IV by proton homonuclear Overhauser enhancement connectivities. The secondary structure in helix IV of the prokaryotic loop is completely demonstrated spectroscopically for the first time in any native or enzyme-cleaved 5S rRNA. In addition, G{sub 21}{center dot}C{sub 58}-A{sub 20}{center dot}U{sub 59}-G{sub 19}{center dot}C{sub 60}-A{sub 18}{center dot}U{sub 61} in helix II, U{sub 32}{center dot}A{sub 46}-G{sub 31}{center dot}C{sub 47}-C{sub 30}{center dot}G{sub 48}-C{sub 29}{center dot}G{sub 49} in helix III, and G{sub 4}{center dot}C{sub 112}-G{sub 5}{center dot}C{sub 111}-U{sub 6}{center dot}G{sub 110} in the terminal stem (helix I) have been assigned by means of NOE experiments on intact 5S rRNA and its fragments A and C. Base pairs in helices I-IV of the universal secondary structure of B. megaterium 5S RNA are described.

• OSTI ID: 6717768
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:3; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
BACILLUS
BACILLUS MEGATERIUM
BACTERIA
BIOCHEMISTRY
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHEMISTRY
DECOMPOSITION
ENZYMATIC HYDROLYSIS
ENZYMES
ESTERASES
HYDROLASES
HYDROLYSIS
LYSIS
MAGNETIC RESONANCE
MICROORGANISMS
MOLECULAR STRUCTURE
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEASES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANOIDS
OVERHAUSER EFFECT
PHOSPHODIESTERASES
RESONANCE
RIBOSOMAL RNA
RIBOSOMES
RNA
SOLVOLYSIS
SPECTRA
TEMPERATURE EFFECTS