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Record W4247569210 · doi:10.24124/2010/bpgub647

Dissection of U4 snRNA functional domains using in vitro reconstitution.

2010· dissertation· en· W4247569210 on OpenAlexaff
Amy Hayduk

Bibliographic record

Venuenot available
Typedissertation
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicRNA Research and Splicing
Canadian institutionsLibrary and Archives Canada
Fundersnot available
KeywordssnRNPSpliceosomePrp24RNA splicingSmall nuclear RNARibonucleoproteinBiologyCell biologySmall nuclear ribonucleoproteinMinor spliceosomeIntronGeneticsSplicing factorComputational biologyRNAGeneNon-coding RNA

Abstract

fetched live from OpenAlex

Splicing, the removal of non-protein-coding introns from pre-mRNA, is a critical step in eukaryotic gene expression, yet this process is still far from being fully understood. Investigation of the splicing mechanism has been hindered by its complexity. The spliceosome, the dynamic ribonucleoprotein complex which catalyzes splicing, contains over 100 molecular components, some of which are organized into small nuclear ribonucleoprotein particles (snRNPs). Each snRNP is composed of a small nuclear RNA (snRNA U1, U2, U4, U5, or U6) and its associated proteins. While the general roles of U1, U2, U5, and U6 are reasonably well understood, the function of U4, though essential for splicing, remains much less clear. It has been demonstrated that U4 forms an extensive base-pairing interaction with U6, but the function and mechanism of formation of this interaction have not been established. One reasonable hypothesis is that interaction of U4 and U6 allows U4 to activate U6 for assembly onto the pre-mRNA. Furthermore, there is only a limited knowledge of the proteins associated with U4, where they bind, and what functions they may perform during snRNP and spliceosome assembly and splicing catalysis. Determination of the structure of the U4 snRNP and its role is splicing has been hindered by lack of an in vitro system allowing U4 reconstitution analyses. To facilitate molecular dissection of the structure and function of U4 in splicing, I have developed an in vitro assay for reconstitution of functional U4 snRNPs. Endogenous U4 was depleted from yeast splicing extract through oligonucleotide-directed RNase H degradation, and optimal conditions for this reaction were identified. The most important factor for efficient U4 degradation was to perform the reaction under conditions of active pre-mRNA splicing, as the splicing process was found to increase accessibility of the targeted region of U4. Depletion of U4 resulted in an early block in spliceosome assembly, strongly inhibiting splicing activity. Subsequent addition of

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.002
Threshold uncertainty score0.009

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0010.000
Meta-epidemiology (narrow)0.0010.001
Meta-epidemiology (broad)0.0000.001
Bibliometrics0.0000.000
Science and technology studies0.0010.000
Scholarly communication0.0010.000
Open science0.0010.001
Research integrity0.0010.002
Insufficient payload (model declined to judge)0.0010.001

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.015
GPT teacher head0.293
Teacher spread0.278 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations0
Published2010
Admission routes1
Has abstractyes

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