Role of cytoskeleton in dynamics of ntp303 mRNA distribution, storage and activation in the process of tobacco pollen maturation and tube growth
Project goals
Tobacco pollen specific ntp303 gene represents a natural model of translational regulation of gene expression. Pollen tube growth is dependent on intensive synthesis of the pollen tube wall glycoprotein, p69, and corresponding ntp303 transcripts activated from stored mRNAs synthesized during pollen maturation. Our pilot study proved that ntp303 transcripts in their repressed form were accumulated in EPPs, EDTA-puromycin resistant particles, and a 30 kDa protein was identified as a candidate for a ntp303mRNA binding protein regulating its translatability (GA AV A5038801 grant, Honys et al. 2000, Honys 2000). Our aim is to study the role of the cytoskeleton in distribution of repressed ntp303 mRNA in EPP particles and its activation and te-localizationinto polysomes. Biochemical techniques will be combined with in situ hybridization, cytoskeleton immunofluorescence and in vitro binding assays to specify the interactions between ntp303 mRNA, RNA-binding proteins and cytoskeleton.
Keywords
Nicotiana tabacum - L. pollenpollen tubestranslational regulation of gene expressionmRNA-binding proteinscytoskeletoncytoskeleton-binding proteinsmRNA translatability
Public support
Provider
Academy of Sciences of the Czech Republic
Programme
Grants of distinctly investigative character focused on the sphere of research pursued at present particularly in the Academy of Sciences of the Czech Republic
Call for proposals
Výzkumné granty 2 (SAV02002-AB)
Main participants
Ústav experimentální botaniky AV ČR, v. v. i.
Contest type
VS - Public tender
Contract ID
—
Alternative language
Project name in Czech
Úloha cytoskeletu v distribuci, skladování a aktivaci ntp 303 mRNA v procesu zrání pylových zrn a růstu pylových láček tabáku
Annotation in Czech
Pylově specifický gen tabáku ntp303 je modelovým příkladem translační regulace genové exprese. Růst pylových láček je podmiňován intensivní syntézou proteinu 69 kDa, jež je součástí stěny pylové láčky. Kodující transkript ntp303 je syntetizován již běhemzrání pylu, je skladován a v pylových láčkách aktivován. Naše analýzy ukázaly, že ntp303 mRNA je v reprimovaném stavu skladována v EPP, EDTA-puromycin resistentních partikulích a translatabilita by mohla být regulována bílkovinou 30 kDa vážící se na ntp303 mRNA (GA AV A5038801, Honys et al. 2000, Honys 2000). Cílem projektu je popsat podíl cytoskeletu na uložení reprimovaných molekul ntp303 mRNP do EPP partikulí a na jejich aktivaci a přemístění na polysomy. Interakce mezi ntp303 mRNA, ntp303 mRNA vazebnými bílkovinami, cytoskeletem a na cytoskelet navázanými bílkovinami budou analyzovány biochemickými i molekulárně biologickými technikami spolu s in situ hybdidizací, zviditelněním cytoskelet a in vitro vazebnými testy.
Scientific branches
R&D category
ZV - Basic research
CEP classification - main branch
EB - Genetics and molecular biology
CEP - secondary branch
EF - Botany
CEP - another secondary branch
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10603 - Genetics and heredity (medical genetics to be 3)
10604 - Reproductive biology (medical aspects to be 3)
10605 - Developmental biology
10608 - Biochemistry and molecular biology
10609 - Biochemical research methods
10611 - Plant sciences, botany
10612 - Mycology
30101 - Human genetics
Completed project evaluation
Provider evaluation
U - Uspěl podle zadání (s publikovanými či patentovanými výsledky atd.)
Project results evaluation
During pollen maturation are formed EPP, supramolecular complexes of mRNPs, proteins and ribosomes, which are connected with actin. Actin and tubulin bound to EPP facilitate their transport and continuous activation of stored mRNPs in tube tips.
Solution timeline
Realization period - beginning
Jan 1, 2002
Realization period - end
Jan 1, 2005
Project status
U - Finished project
Latest support payment
—
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP06-AV0-IA-U/04:2
Data delivery date
Sep 26, 2007
Finance
Total approved costs
10,105 thou. CZK
Public financial support
3,254 thou. CZK
Other public sources
4,649 thou. CZK
Non public and foreign sources
2,202 thou. CZK
Recognised costs
10 105 CZK thou.
Public support
3 254 CZK thou.
0%
Provider
Academy of Sciences of the Czech Republic
CEP
EB - Genetics and molecular biology
Solution period
01. 01. 2002 - 01. 01. 2005