tRNA editing by deamination: Balancing affinity and specificity
tRNA editing by deamination: Balancing affinity and specificity
批准号:
10389330
负责人:
Juan D Alfonzo
金额:
$7.23万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2008
资助国家:
美国
项目状态:
已结题
起止时间:
2008-08-01 至 2023-11-30
关键词:
12 year oldAffectAffinityAnabolismAnticodonArginine Specific tRNABindingBiochemicalBiological AssayCatalysisCell NucleusCellsChemicalsCodon NucleotidesCytosineDeaminaseDeaminationDeuteriumElectrophoretic Mobility Shift AssayEnzymesEquilibriumEquipmentEukaryotaFundingGene ExpressionGenetic TranscriptionGenomeGenomicsHumanIn VitroIndividualLabelLaboratoriesLeishmaniaLuciferasesMapsMass Spectrum AnalysisMediatingMedicalMethylationMethyltransferaseModelingModificationMolecular AnalysisMutagenesisNational Institute of General Medical SciencesNuclearNucleic AcidsOhioOrganismPathway interactionsPositioning AttributeProteinsProteomicsPurinesPyrimidineRNA Recognition MotifRadioactiveReactionRegulationReporterResearchRibosomesServicesSiteSpecific qualifier valueSpecificitySystemTechniquesTestingThreonine-Specific tRNATransfer RNATranslationsTrypanosomaTrypanosoma brucei bruceiUniversitiesUntranslated RegionsUridineVariantVisualizationbaseenzyme activityin vitro activityin vivoinstrumentparent grantreconstitutionribosome profilingsugartranscriptome
中文摘要
核酸经过自然发生的化学修饰。超过100种不同的
英文摘要
Nucleic acids undergo naturally occurring chemical modifications. Over 100 different
modifications have been described and every position in the purine and pyrimidine
bases can be modified; often the sugar is also modified. Despite recent progress, the
mechanism for the biosynthesis of most modifications is not fully understood, owing,
in part, to the difficulty associated with reconstituting enzyme activity in vitro. Whereas
some modifications can be efficiently formed with purified components, others may
require more intricate pathways. A model for modification interdependence, in which
one modification is a prerequisite for another, potentially explains a major hindrance
in reconstituting enzymatic activity in vitro. This model was prompted by our earlier
discovery of tRNA cytosine-to-uridine editing in eukaryotes; a reaction that had not
been recapitulated in vitro and the mechanism for which remains unknown. Formation
of m3C in vitro requires the presence of both, the T. brucei methyltransferase TRM140
and the deaminase ADAT2/3. Once formed m3C is deaminated to m3U by the same
set of enzymes.
The propose research relies heavily on our ability to determine the specific
contributions of individual residues and domains of TRM140 and ADAT2/3 to
substrate recognition and catalysis. Binding in my laboratory is determined by
Eletrophoretic Mobility Shift Assays (EMSA), while determination of catalytic activity
relies on either deamination of methylation assays based on incubation a
radioactively labeled substrate with the enzyme(s) in question. The results of both
assays are forcibly visualized and quantified by using a Typhoon-type
PhosphorImager system. The current supplement is to replace an existing unit which
has broken down and it is no longer serviceable. Without it, successful completion of
the research proposed is nearly impossible
This request is being submitted in parallel with a proposal from Dr. Kurt Fredrick in
my department at Ohio State University who also requires this equipment for his
project entitled “Molecular analysis of accurate ribosomal translocation” (R01
GM072528).
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DOI:
10.3390/life6010013
发表时间:
2016-03-14
期刊:
Life (Basel, Switzerland)
影响因子:
--
作者:
[McKenney KM, Alfonzo JD]
通讯作者:
Alfonzo JD
RNAi, the guiding principle and keeping family happy.
RNAi,指导原则和保持家庭幸福。
DOI:
10.1261/rna.049635.115
发表时间:
2015
期刊:
RNA (New York, N.Y.)
影响因子:
--
作者:
[Alfonzo,JuanD]
通讯作者:
Alfonzo,JuanD
DOI:
10.1002/iub.1957
发表时间:
2018-12
期刊:
IUBMB life
影响因子:
4.6
作者:
[Paris Z, Alfonzo JD]
通讯作者:
Alfonzo JD
DOI:
10.1261/rna.062893.117
发表时间:
2018-01
期刊:
RNA (New York, N.Y.)
影响因子:
--
作者:
[McKenney KM, Rubio MAT, Alfonzo JD]
通讯作者:
Alfonzo JD
DOI:
10.1016/j.molcel.2013.08.042
发表时间:
2013-10-24
期刊:
MOLECULAR CELL
影响因子:
16
作者:
[Rubio, Mary Anne T., Paris, Zdenek, Gaston, Kirk W., Fleming, Ian M. C., Sample, Paul, Trotta, Christopher R., Alfonzo, Juan D.]
通讯作者:
Alfonzo, Juan D.
共 23 条
Study of queuosine salvage and function in eukaryotes; a forgotten micronutrient
-
批准号:10080744
-
项目类别:
-
资助金额:$38.29万
-
财政年份:2019
-
负责人:Juan D Alfonzo
-
依托单位:
Study of queuosine salvage and function in eukaryotes; a forgotten micronutrient
-
批准号:10319932
-
项目类别:
-
资助金额:$38.29万
-
财政年份:2019
-
负责人:Juan D Alfonzo
-
依托单位:
Study of queuosine salvage and function in eukaryotes; a forgotten micronutrient
-
批准号:9904725
-
项目类别:
-
资助金额:$38.29万
-
财政年份:2019
-
负责人:Juan D Alfonzo
-
依托单位:
The Mechanism of tRNA splicing in trypanosomes
-
批准号:9531616
-
项目类别:
-
资助金额:$43.89万
-
财政年份:2017
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificity
-
批准号:7532281
-
项目类别:
-
资助金额:$31.94万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificity
-
批准号:9767224
-
项目类别:
-
资助金额:$36.0万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificty
-
批准号:8858638
-
项目类别:
-
资助金额:$31.57万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificity
-
批准号:8074072
-
项目类别:
-
资助金额:$30.12万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificity
-
批准号:7662426
-
项目类别:
-
资助金额:$30.73万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificty
-
批准号:8479370
-
项目类别:
-
资助金额:$30.46万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificty
-
批准号:8321155
-
项目类别:
-
资助金额:$32.88万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificty
-
批准号:8665968
-
项目类别:
-
资助金额:$31.57万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
tRNA editing by deamination: Balancing affinity and specificity
-
批准号:7821385
-
项目类别:
-
资助金额:$30.43万
-
财政年份:2008
-
负责人:Juan D Alfonzo
-
依托单位:
海外基金