Limitation of Ribosomal Protein L11 Availability in vivo Affects Translation Termination

Dyke, Natalya Van; Xu, Wenjian; Murgola, Emanuel J.

doi:10.1016/s0022-2836(02)00304-2

Cited by 27 publications

(31 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…19 Similarly, the RF3-dependent release of RF1 and RF2 is drastically decreased when the uL11 protein is absent on the ribosome during termination. 20 In the case of eukaryotic uL11, its structural and functional mode of action is still less understood; however, its structure was determined on eukaryotic ribosomes, and interaction between yeast uL11 protein and eEF2 was postulated. 21 So far, there are scattered biochemical data showing that the absence of the uL11 protein impairs the translation elongation step, in both efficiency and accuracy meanings.…”

Section: Introductionmentioning

confidence: 99%

Functional analysis of the uL11 protein impact on translational machinery

Wawiórka¹,

Molestak²,

Szajwaj³

et al. 2016

Cell Cycle

View full text Add to dashboard Cite

The ribosomal GTPase associated center constitutes the ribosomal area, which is the landing platform for translational GTPases and stimulates their hydrolytic activity. The ribosomal stalk represents a landmark structure in this center, and in eukaryotes is composed of uL11, uL10 and P1/P2 proteins. The modus operandi of the uL11 protein has not been exhaustively studied in vivo neither in prokaryotic nor in eukaryotic cells. Using a yeast model, we have brought functional insight into the translational apparatus deprived of uL11, filling the gap between structural and biochemical studies. We show that the uL11 is an important element in various aspects of 'ribosomal life'. uL11 is involved in 'birth' (biogenesis and initiation), by taking part in Tif6 release and contributing to ribosomal subunit-joining at the initiation step of translation. uL11 is particularly engaged in the 'active life' of the ribosome, in elongation, being responsible for the interplay with eEF1A and fidelity of translation and contributing to a lesser extent to eEF2-dependent translocation. Our results define the uL11 protein as a critical GAC element universally involved in trGTPase 'productive state' stabilization, being primarily a part of the ribosomal element allosterically contributing to the fidelity of the decoding event.

show abstract

Section: Introductionmentioning

confidence: 99%

Functional analysis of the uL11 protein impact on translational machinery

Wawiórka¹,

Molestak²,

Szajwaj³

et al. 2016

Cell Cycle

View full text Add to dashboard Cite

show abstract

“…Deletion of the distal part of this b-hairpin in a-sarcin (residues 7-22) resulted in a D (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22) mutant that had lost the ability to cleave specifically the ribosome but was still an active ribonuclease able to specifically hydrolyze a SRL-like 35-mer oligoribonucleotide [35]. Its three-dimensional structure in solution [36] showed that the folding of wild-type a-sarcin was preserved, including the spatial conformation of the loops (Fig.…”

Section: Introductionmentioning

confidence: 99%

Modeling the highly specific ribotoxin recognition of ribosomes

García-Mayoral¹,

Garcı́a-Ortega²,

Álvarez-García³

et al. 2005

FEBS Letters

View full text Add to dashboard Cite

The three-dimensional structures of the a-sarcin ribotoxin and its D(7-22) deletion mutant, both complexed with a 20-mer oligonucleotide mimicking the sarcin/ricin loop (SRL) of the ribosome, have been docked into the structure of the Halobacterium marismortui ribosome by fitting the nucleotide atomic coordinates into those of the ribosomal SRL. This study has revealed that two regions of the ribotoxin, residues 11-16 and 84-85, contact the ribosomal proteins L14 (residues 99-105) and L6 (residues 88-92), respectively. The first of these two ribotoxin regions appears to be crucial for its specific ribosome recognition.

show abstract

“…Bacterial Strains and Plasmids-The three different E. coli strains used for ribosome preparation (6,18) (Table 1) carry a chromosomal knock-out of the rplK gene, which encodes the L11 protein. In the FTP6063 and FTP6066 strains the knock-outs were complemented with the whole L11 gene or its CTD cloned in the p⌬CAT plasmid, which are referred to as pL11 and pL11Cter, respectively (18).…”

Section: Components Of the In Vitro Translation Systemmentioning

confidence: 99%

“…L11 binds to the nucleotides 1051-1108 of Escherichia coli 23 S rRNA, commonly called the L11 binding region (L11BR) 2 (6), which constitutes the GTPase-associated center, an important sector of the bacterial ribosome, where all of the translational GTPases bind and hydrolyze GTP in the course of their action (7). This is also the site of action for the thiazole antibiotics thiostrepton and micrococcin (8 -10).…”

mentioning

confidence: 99%

“…The CTD of L11 is in stable contact with the L11BR RNA, whereas the NTD can change its position and proximity with respect to the rest of the protein (19). Earlier biochemical and genetic studies indicate that depletion of L11 from the ribosome inhibits RF1-mediated translation termination on a UAG codon but somewhat facilitates RF2-mediated termination on a UGA codon (4,6). Antibodies against the NTD of L11 inhibit RF1-mediated termination, indicating that this part of L11 is crucial for RF1 binding (14).…”

mentioning

confidence: 99%

See 1 more Smart Citation

The Role of Ribosomal Protein L11 in Class I Release Factor-mediated Translation Termination and Translational Accuracy

Bouakaz

Murgola

et al. 2006

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

It has been suggested from in vivo and cryoelectron micrographic studies that the large ribosomal subunit protein L11 and its N-terminal domain play an important role in peptide release by, in particular, the class I release factor RF1. In this work, we have studied in vitro the role of L11 in translation termination with ribosomes from a wild type strain (WT-L11), an L11 knocked-out strain (⌬L11), and an L11 N terminus truncated strain (Cter-L11). Our data show 4 -6-fold reductions in termination efficiency (k cat /K m ) of RF1, but not of RF2, on ⌬L11 and Cter-L11 ribosomes compared with wild type. There is, at the same time, no effect of these L11 alterations on the maximal rate of ester bond cleavage by either RF1 or RF2. The rates of dissociation of RF2 but not of RF1 from the ribosome after peptide release are somewhat reduced by the L11 changes irrespective of the presence of RF3, and they cause a 2-fold decrease in the missense error. Our results suggest that the L11 modifications increase nonsense suppression at UAG codons because of the reduced termination efficiency of RF1 and that they decrease nonsense suppression at UGA codons because of a decreased missense error level.L11 is a highly conserved ribosomal 14.8-kDa protein located at the base of the L7/L12 stalk of the ribosome, which is essential for several steps in protein synthesis (1-5). L11 binds to the nucleotides 1051-1108 of Escherichia coli 23 S rRNA, commonly called the L11 binding region (L11BR) 2 (6), which constitutes the GTPase-associated center, an important sector of the bacterial ribosome, where all of the translational GTPases bind and hydrolyze GTP in the course of their action (7). This is also the site of action for the thiazole antibiotics thiostrepton and micrococcin (8 -10). In addition to the GTPases, some other translational factors interact with L11 and the L11BR in functionally important ways. The class I release factors RF1 and RF2 belong to this group. RF1 recognizes the stop codons UAG and UAA, whereas RF2 recognizes the stop codons UGA and UAA in the A site of the ribosome (11). Recent cryo-EM studies with termination complexes containing RF2 (12, 13) and RF1 3 show that these two factors acquire very similar overall conformations on the ribosome. Although they bind to the decoding center on the 30 S subunit and reach up to the peptidyltransferase center on the 50 S subunit to induce release of the nascent peptide chain, they interact with L11BR (12, 13) and L11 3 with their flexible domain I. These observations are in line with previous suggestions, based on biochemical and genetic experiments, that there are interactions between L11 and the release factors (4, 14 -18).The L11 protein consists of two domains, a tightly folded N-terminal domain (NTD), which is loosely connected to the large compact C-terminal domain (CTD). The CTD of L11 is in stable contact with the L11BR RNA, whereas the NTD can change its position and proximity with respect to the rest of the protein (19). Earlier biochemical and genetic studies i...

show abstract

Limitation of Ribosomal Protein L11 Availability in vivo Affects Translation Termination

Cited by 27 publications

References 35 publications

Functional analysis of the uL11 protein impact on translational machinery

Functional analysis of the uL11 protein impact on translational machinery

Modeling the highly specific ribotoxin recognition of ribosomes

The Role of Ribosomal Protein L11 in Class I Release Factor-mediated Translation Termination and Translational Accuracy

Contact Info

Product

Resources

About