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Tytuł pozycji:

Translational regulation of human neuronal nitric-oxide synthase by an alternatively spliced 5'-untranslated region leader exon.

Tytuł:
Translational regulation of human neuronal nitric-oxide synthase by an alternatively spliced 5'-untranslated region leader exon.
Autorzy:
Newton DC; Renal Division and the Department of Medicine, St. Michael's Hospital and University of Toronto, Toronto, Ontario M5S 1A8, Canada.
Bevan SC
Choi S
Robb GB
Millar A
Wang Y
Marsden PA
Źródło:
The Journal of biological chemistry [J Biol Chem] 2003 Jan 03; Vol. 278 (1), pp. 636-44. Date of Electronic Publication: 2002 Oct 25.
Typ publikacji:
Journal Article; Research Support, Non-U.S. Gov't
Język:
English
Imprint Name(s):
Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
MeSH Terms:
Alternative Splicing*
Gene Expression Regulation, Enzymologic*
Protein Biosynthesis*
5' Untranslated Regions/*genetics
Nitric Oxide Synthase/*genetics
5' Untranslated Regions/metabolism ; Animals ; Base Sequence ; Cell Line ; Exons/genetics ; Genes, Reporter ; Humans ; Mice ; Molecular Sequence Data ; Nitric Oxide Synthase/metabolism ; Nitric Oxide Synthase Type I ; Nucleic Acid Conformation ; Open Reading Frames ; Promoter Regions, Genetic ; RNA Caps ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Rats ; Tissue Distribution
Molecular Sequence:
GENBANK AY098642
Substance Nomenclature:
0 (5' Untranslated Regions)
0 (RNA Caps)
0 (RNA, Messenger)
EC 1.14.13.39 (NOS1 protein, human)
EC 1.14.13.39 (Nitric Oxide Synthase)
EC 1.14.13.39 (Nitric Oxide Synthase Type I)
EC 1.14.13.39 (Nos1 protein, mouse)
EC 1.14.13.39 (Nos1 protein, rat)
Entry Date(s):
Date Created: 20021031 Date Completed: 20030210 Latest Revision: 20210209
Update Code:
20240104
DOI:
10.1074/jbc.M209988200
PMID:
12403769
Czasopismo naukowe
Expression of the neuronal nitric-oxide synthase (nNOS) mRNA is subject to complex cell-specific transcriptional regulation, which is mediated by alternative promoters. Unexpectedly, we identified a 89-nucleotide alternatively spliced exon located in the 5'-untranslated region between exon 1 variants and a common exon 2 that contains the translational initiation codon. Alternative splicing events that do not affect the open reading frame are distinctly uncommon in mammals; therefore, we assessed its functional relevance. Transient transfection of reporter RNAs performed in a variety of cell types revealed that this alternatively spliced exon acts as a potent translational repressor. Stably transfected cell lines confirmed that the alternatively spliced exon inhibited translation of the native nNOS open reading frame. Reverse transcription-PCR and RNase protection assays indicated that nNOS mRNAs containing this exon are common and expressed in both a promoter-specific and tissue-restricted fashion. Mutational analysis identified the functional cis-element within this novel exon, and a secondary structure prediction revealed that it forms a putative stem-loop. RNA electrophoretic mobility shift assay techniques revealed that a specific cytoplasmic RNA-binding complex interacts with this motif. Hence, a unique splicing event within a 5'-untranslated region is demonstrated to introduce a translational control element. This represents a newer model for the translational control of a mammalian mRNA.

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