Functional analysis of the uL11 protein impact on translational machinery

Wawiórka, Leszek; Molestak, Eliza; Szajwaj, Monika; Michalec-Wawiórka, Barbara; Boguszewska, Aleksandra; Borkiewicz, Lidia; Liudkovska, Vladyslava; Kufel, Joanna; Tchórzewski, Marek

doi:10.1080/15384101.2016.1154245

Cited by 15 publications

(29 citation statements)

References 69 publications

(86 reference statements)

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“…variants. Our findings also corroborate observations in yeast that RPL12 deletion slows translation (11,12) and complement these data with the discovery that the overall effect is global, albeit dependent on the specific protein defect and/or codon context. For what we believe is the first time, we used this approach to establish impact of disease-relevant mutations on CFTR biogenesis from a mechanistic perspective, including effects on elongation.…”

Section: Figure 3 Silencing Rpl12 Enhances F508del-cftr Functional Esupporting

confidence: 90%

Slowing ribosome velocity restores folding and function of mutant CFTR

Oliver¹,

Rauscher²,

Mijnders³

et al. 2019

Journal of Clinical Investigation

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Section: Figure 3 Silencing Rpl12 Enhances F508del-cftr Functional Esupporting

confidence: 90%

Slowing ribosome velocity restores folding and function of mutant CFTR

Oliver¹,

Rauscher²,

Mijnders³

et al. 2019

Journal of Clinical Investigation

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“…In such a situation, EF-2-catalyzed translocation should be affected to the same extent as decoding, because elongation consumes equal numbers of eEF-2-and eEF-1A-dependent steps. Interestingly, such a situation was indeed described for uL11-depleted yeast cells (64). Since uL11 is present on the ribosome in a single copy, ribosomes lacking this protein were equally defective in all trGTPase dependent steps, including ribosome biogenesis, subunit joining, translation elongation, and termination, and especially affecting central ribosomal activity, the elongation cycle, which was depicted as perturbations in misincorporation, and Ϫ1 and ϩ1 PRF (64).…”

Section: P1/p2 Proteins Are Dispensable For the Translational Speed Imentioning

confidence: 80%

Multiplication of Ribosomal P-Stalk Proteins Contributes to the Fidelity of Translation

Wawiórka

Molestak

Szajwaj

et al. 2017

Molecular and Cellular Biology

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The P-stalk represents a vital element within the ribosomal GTPaseassociated center, which represents a landing platform for translational GTPases. The eukaryotic P-stalk exists as a uL10-(P1-P2) 2 pentameric complex, which contains five identical C-terminal domains, one within each protein, and the presence of only one such element is sufficient to stimulate factor-dependent GTP hydrolysis in vitro and to sustain cell viability. The functional contribution of the P-stalk to the performance of the translational machinery in vivo, especially the role of P-protein multiplication, has never been explored. Here, we show that ribosomes depleted of P1/P2 proteins exhibit reduced translation fidelity at elongation and termination steps. The elevated rate of the decoding error is inversely correlated with the number of the P-proteins present on the ribosome. Unexpectedly, the lack of P1/P2 has little effect in vivo on the efficiency of other translational GTPase (trGTPase)-dependent steps of protein synthesis, including translocation. We have shown that loss of accuracy of decoding caused by P1/P2 depletion is the major cause of translation slowdown, which in turn affects the metabolic fitness of the yeast cell. We postulate that the multiplication of P-proteins is functionally coupled with the qualitative aspect of ribosome action, i.e., the recoding phenomenon shaping the cellular proteome.KEYWORDS ribosomal proteins, ribosomal stalk, ribosome A t the expense of energy from GTP hydrolysis, translational GTPases (trGTPases) confer the unidirectional trajectory for the translational apparatus, providing at the same time unique timing for individual steps (1, 2). The main landing platform for trGTPases is situated on the large ribosomal subunit called the GTPase-associated center (GAC), and it represents a universally conserved ribosomal element where stimulation of trGTPase catalytic activity takes place (3). The GAC consists of two main elements, a conserved fragment of rRNA called the sarcin-ricin loop (SRL) and a ribosomal stalk composed of ribosomal proteins, which form an oligomeric protein complex (4). The protein part of GAC, the ribosome stalk, can be divided into two functionally and evolutionarily distinct parts, the base of the stalk and its lateral elements. The stalk base is composed of conserved ribosomal proteins uL11 (former names L11 and L12 for prokaryotes and eukaryotes, respectively) and uL10 (former names L10 and P0), which anchor the stalk to the rRNA (5, 6). The lateral part of the stalk is built of dimeric complexes P1-P2 in eukaryotes/archaea or (bL12) 2 in prokaryotes (4, 7). Despite the lack of amino acid sequence conservation, the lateral stalk fulfils the same functions and has a similar structural architecture across all domains of life (8). P1/P2 and bL12 proteins are built of two domains. The globular N-terminal domain (NTD) is responsible for dimerization, whereas the highly acidic C-terminal domain (CTD) interacts with trGTPases (9, 10). However, the structure of the CTD in euk...

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“…It has been shown that the stable association of Yvh1 with pre-60S r-particles depends on the r-protein L12 (uL11), which is the closest neighbour of P0 at the base of the P-stalk 4447. However, still in the absence of L12, there is apparently no difference in the efficiency of Mrt4 re-importation to the nucleus (48, and our unpublished results). In conclusion, despite the fact that yeast Mrt4 and Yvh1 are non-essential proteins under standard laboratory conditions, both factors may play important roles controlling the position and timing of the assembly of P0, simultaneously providing a surveillance point to ensure that only mature LSUs can engage in translation (see below; further discussed in 141720).…”

Section: The Classical View: Paralogues Of Ribosomal Proteinsmentioning

confidence: 67%

Placeholder factors in ribosome biogenesis: please, pave my way

Espinar-Marchena¹,

Babiano²,

Cruz³

2017

Microb Cell

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The synthesis of cytoplasmic eukaryotic ribosomes is an extraordinarily energy-demanding cellular activity that occurs progressively from the nucleolus to the cytoplasm. In the nucleolus, precursor rRNAs associate with a myriad of trans-acting factors and some ribosomal proteins to form pre-ribosomal particles. These factors include snoRNPs, nucleases, ATPases, GTPases, RNA helicases, and a vast list of proteins with no predicted enzymatic activity. Their coordinate activity orchestrates in a spatiotemporal manner the modification and processing of precursor rRNAs, the rearrangement reactions required for the formation of productive RNA folding intermediates, the ordered assembly of the ribosomal proteins, and the export of pre-ribosomal particles to the cytoplasm; thus, providing speed, directionality and accuracy to the overall process of formation of translation-competent ribosomes. Here, we review a particular class of trans-acting factors known as "placeholders". Placeholder factors temporarily bind selected ribosomal sites until these have achieved a structural context that is appropriate for exchanging the placeholder with another site-specific binding factor. By this strategy, placeholders sterically prevent premature recruitment of subsequently binding factors, premature formation of structures, avoid possible folding traps, and act as molecular clocks that supervise the correct progression of pre-ribosomal particles into functional ribosomal subunits. We summarize the current understanding of those factors that delay the assembly of distinct ribosomal proteins or subsequently bind key sites in pre-ribosomal particles. We also discuss recurrent examples of RNA-protein and protein-protein mimicry between rRNAs and/or factors, which have clear functional implications for the ribosome biogenesis pathway.

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Functional analysis of the uL11 protein impact on translational machinery

Cited by 15 publications

References 69 publications

Slowing ribosome velocity restores folding and function of mutant CFTR

Slowing ribosome velocity restores folding and function of mutant CFTR

Multiplication of Ribosomal P-Stalk Proteins Contributes to the Fidelity of Translation

Placeholder factors in ribosome biogenesis: please, pave my way

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