The use of azidoarylimidoesters in RNA-protein cross-linking studies with Escherichia coli ribosomes

Rinke, Jutta; Meinke, Marlis; Brimacombe, Richard; Fink, Gerald R.; Rommel, W.; Fasold, H.

doi:10.1016/0022-2836(80)90318-6

Cited by 30 publications

(18 citation statements)

References 23 publications

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“…30S and 50S ribosomal subunits from E. coli MRE 600 were prepared as previously described (13,14,6). The subunits were either unlabelled, or labelled with 32P in the RNA moiety, or labelled with 3H in the RNA and 14C in the protein moiety.…”

Section: Methodsmentioning

confidence: 99%

“…(a) To determine the extent of reaction, aliquots of doublelabelled 30S or 50S subunits (1-2 A260 units) were used. After cross-linking as described above, the reaction mixtures were applied to 3-8% polyacrylamide gels containing dodecyl sulphate (1,2,6). The percentage of RNA-protein cross-linking was determined from the amount of 14C-protein co-migrating with the peak of 3H-RNA in the gel.…”

Section: Methodsmentioning

confidence: 99%

“…3 A260 units of double-labelled subunit. After crosslinking, the reaction mixtures were applied to sucrose gradients in dodecyl sulphate (1,2,6). The peaks of RNA plus RNA-protein cross-linked complex isolated from the gradients were in each case subjected to digestion with ribonucleases A and T1 as previously described (2).…”

Section: Methodsmentioning

confidence: 99%

“…Our objective has been to introduce such cross-links in situ into the ribosomal subunits, and then to determine the precise sites of cross-linking on both protein and RNA, in order to build up a detailed topographical map of RNA-protein contacts and neighbourhoods within the particles. Up to now this research has been pursued in two directions, firstly the search for suitable bifunctional cross-linking reagents (1,2,4,6) and secondly the development of techniques for the analysis of the cross-link sites (3,5,7).…”

Section: Introductionmentioning

confidence: 99%

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The use of 2-iminothiolane as an RNA-protein cross-linking agent in Escherichia coli ribosomes, and the localisation on 23S RNA of sites cross-linked to proteins L4, L6, L21, L23, L27 and L29

Wower

Meinke

et al. 1981

Nucl Acids Res

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When E. coli ribosomal subunits are reacted with 2-iminothiolane and then subjected to a mild ultraviolet irradiation, an RNA-protein cross-linking reaction occurs. About 5% of the total protein in each subunit becomes cross-linked to the RNA, and a specific sub-set of proteins is involved in the reaction. In the case of the 50S subunit, the sites of cross-linking to the 23S RNA have been determined for six of these proteins: protein L4 is cross-linked within an oligonucleotide comprising positions 613-617 in the 23S sequence, L6 within positions 2473-2481, L21 within positions 540-548, L23 within positions 137-141, L27 within positions 2332-2337 and L29 within positions 99-107.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The use of 2-iminothiolane as an RNA-protein cross-linking agent in Escherichia coli ribosomes, and the localisation on 23S RNA of sites cross-linked to proteins L4, L6, L21, L23, L27 and L29

Wower

Meinke

et al. 1981

Nucl Acids Res

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“…A salient question that arises when studying the crosslinking of a given protein in a particle as complex as the ribosome is whether the considered pro- Turschinsky et af., 1978); 366 nm (s'U) (this paper) and EDC (Chiaruttini ef al., 1980-82) or by means of a bridge with iminothiolane (Wover ef al., 1981-83), bisketoxal (Brewer et al, 1983a,b) and p-azido imido esters (Rinke et al, 1980;. The detection methods for analysis of the proteins are indicated.…”

Section: Rna-protein Crosslinking In Thiolated Ribosomal Subunitsmentioning

confidence: 99%

RNA PROTEIN CROSSLINKS INTRODUCED INTO E. coli RIBOSOMES BY USE OF THE INTRINSIC PROBE 4‐THIOURIDINE

Hajnsdorf

Dubreuil

Bezerra

et al. 1987

Photochem & Photobiology

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Abstract— 70S Ribosome substituted by the uridine photoactivable analogue 4‐thiouridine has been prepared by an in vivo method (substitution level 4.5%). The r‐proteins crosslinked to 16S and 23S rRNA before and after 366‐nm photoactivation were identified. Proteins S2‐S7‐S9/11‐S18 are found linked to 16S RNA in dark‐prepared 30S subunits. Illumination increases uniformly their binding by a factor of 2.5. Similarly, proteins L5‐L15‐L18‐L23‐L28‐L32 are found crosslinked to 23S RNA in dark‐prepared 50S subunits. Photoactivation increases their binding but in addition promotes the covalent linking of proteins L1‐L3‐L4.

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Relevance of Adhesion in Fabrication of Microarrays in Clinical Diagnostics

Kant¹,

Bhatt²,

Sundriyal³

et al. 2017

Adhesion in Pharmaceutical, Biomedical and Dental Fields

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The use of azidoarylimidoesters in RNA-protein cross-linking studies with Escherichia coli ribosomes

Cited by 30 publications

References 23 publications

The use of 2-iminothiolane as an RNA-protein cross-linking agent in Escherichia coli ribosomes, and the localisation on 23S RNA of sites cross-linked to proteins L4, L6, L21, L23, L27 and L29

The use of 2-iminothiolane as an RNA-protein cross-linking agent in Escherichia coli ribosomes, and the localisation on 23S RNA of sites cross-linked to proteins L4, L6, L21, L23, L27 and L29

RNA PROTEIN CROSSLINKS INTRODUCED INTO E. coli RIBOSOMES BY USE OF THE INTRINSIC PROBE 4‐THIOURIDINE

Relevance of Adhesion in Fabrication of Microarrays in Clinical Diagnostics

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