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Graifer, D. M.; Карпова, Г. Г.; Knorre, D.G.

doi:10.1023/a:1010220627612

Cited by 15 publications

(8 citation statements)

References 30 publications

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“…This implies that the protein environment of the first nucleotide of the A site bound codon is similar at the elongation and the termination steps of translation. Based on earlier data on crosslinking of human 80S ribosomes to short mRNA analogs bearing various modifying groups at the 5′‐terminal phosphate, protein S2 was suggested to be located in the vicinity of the mRNA sequence between the first nucleotide of the E site bound codon and the first nucleotide of the P site bound codon on the elongating ribosome [18]. Crosslinking of protein S2 to the last modified nucleotide of the mRNA stop signal UAAA (Fig.…”

Section: Discussionmentioning

confidence: 96%

Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes

et al. 2002

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To study positioning of the mRNA stop signal with respect to polypeptide chain release factors (RFs) and ribosomal components within human 80S ribosomes, photoreactive mRNA analogs were applied. Derivatives of the UUCUAAA heptaribonucleotide containing the UUC codon for Phe and the stop signal UAAA, which bore a perfluoroaryl azido group at either the fourth nucleotide or the 3P P-terminal phosphate, were synthesized. The UUC codon was directed to the ribosomal P site by the cognate tRNA Phe , targeting the UAA stop codon to the A site. Mild UV irradiation of the ternary complexes consisting of the 80S ribosome, the mRNA analog and tRNA resulted in tRNA-dependent crosslinking of the mRNA analogs to the 40S ribosomal proteins and the 18S rRNA. mRNA analogs with the photoreactive group at the fourth uridine (the first base of the stop codon) crosslinked mainly to protein S15 (and much less to S2). For the 3P P-modified mRNA analog, the major crosslinking target was protein S2, while protein S15 was much less crosslinked. Crosslinking of eukaryotic (e) RF1 was entirely dependent on the presence of a stop signal in the mRNA analog. eRF3 in the presence of eRF1 did not crosslink, but decreased the yield of eRF1 crosslinking. We conclude that (i) proteins S15 and S2 of the 40S ribosomal subunit are located near the A site-bound codon; (ii) eRF1 can induce spatial rearrangement of the 80S ribosome leading to movement of protein L4 of the 60S ribosomal subunit closer to the codon located at the A site; (iii) within the 80S ribosome, eRF3 in the presence of eRF1 does not contact the stop codon at the A site and is probably located mostly (if not entirely) on the 60S subunit. ß

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Section: Discussionmentioning

confidence: 96%

Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes

et al. 2002

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“…Another conspicuous difference concerns the mRNA tunnel upstream the E site. In 80S ribosomes the mRNA of this region is surrounded by proteins that are not present in prokaryotes, whereas in 70S ribosomes both ribosomal proteins and rRNA are found (11,12). Undoubtedly, discovery and design of new antibacterial drugs need information about organization, functions and evolutionary conservation of mammalian elongation apparatus.…”

Section: Introductionmentioning

confidence: 99%

Features of 80S mammalian ribosome and its subunits

Budkevich

El'skaya

Nierhaus

2008

Nucleic Acids Research

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It is generally believed that basic features of ribosomal functions are universally valid, but a systematic test still stands out for higher eukaryotic 80S ribosomes. Here we report: (i) differences in tRNA and mRNA binding capabilities of eukaryotic and bacterial ribosomes and their subunits. Eukaryotic 40S subunits bind mRNA exclusively in the presence of cognate tRNA, whereas bacterial 30S do bind mRNA already in the absence of tRNA. 80S ribosomes bind mRNA efficiently in the absence of tRNA. In contrast, bacterial 70S interact with mRNA more productively in the presence rather than in the absence of tRNA. (ii) States of initiation (Pi), pre-translocation (PRE) and post-translocation (POST) of the ribosome were checked and no significant functional differences to the prokaryotic counterpart were observed including the reciprocal linkage between A and E sites. (iii) Eukaryotic ribosomes bind tetracycline with an affinity 15 times lower than that of bacterial ribosomes (Kd 30 μM and 1–2 μM, respectively). The drug does not effect enzymatic A-site occupation of 80S ribosomes in contrast to non-enzymatic tRNA binding to the A-site. Both observations explain the relative resistance of eukaryotic ribosomes to this antibiotic.

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“…The first guide sequence (from the 5′-terminus) was directed to G1702 in 18S rRNA, which was one of the key nucleotides of the ribosome-decoding center [25]. Earlier, we studied the changes in the 18S rRNA structure after transfection of artificial box C/D RNAs into human cells.…”

Section: Resultsmentioning

confidence: 99%

Artificial Box C/D RNAs Affect Pre-mRNA Maturation in Human Cells

Степанов

Semenov

Savelyeva

et al. 2013

BioMed Research International

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Box C/D small nucleolar RNAs (snoRNAs) are known to guide the 2′-O-ribose methylation of nucleotides in eukaryotic ribosomal RNAs and small nuclear RNAs. Recently snoRNAs are predicted to regulate posttranscriptional modifications of pre-mRNA. To expand understanding of the role of snoRNAs in control of gene expression, in this study we tested the ability of artificial box C/D RNAs to affect the maturation of target pre-mRNA. We found that transfection of artificial box C/D snoRNA analogues directed to HSPA8 pre-mRNAs into human cells induced suppression of the target mRNA expression in a time- and dose-dependent manner. The artificial box C/D RNA directed to the branch point adenosine of the second intron, as well as the analogue directed to the last nucleotide of the second exon of the HSPA8 pre-mRNA caused the most prominent influence on the level of HSPA8 mRNAs. Neither box D nor the ability to direct 2′-O-methylation of nucleotides in target RNA was essential for the knockdown activity of artificial snoRNAs. Inasmuch as artificial box C/D RNAs decreased viability of transfected human cells, we propose that natural snoRNAs as well as their artificial analogues can influence the maturation of complementary pre-mRNA and can be effective regulators of vital cellular processes.

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Untitled

Cited by 15 publications

References 30 publications

Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes

Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes

Features of 80S mammalian ribosome and its subunits

Artificial Box C/D RNAs Affect Pre-mRNA Maturation in Human Cells

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