The Elav-like Proteins Bind to a Conserved Regulatory Element in the 3′-Untranslated Region of GAP-43 mRNA

Chung, Sangmi; Eckrich, Michael J.; Perrone‐Bizzozero, Nora I.; Kohn, Douglas T.; Furneaux, Henry

doi:10.1074/jbc.272.10.6593

Cited by 141 publications

(142 citation statements)

References 41 publications

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“…HuR (8) is a ubiquitously expressed member of the embryonic lethal abnormal vision (ELAV) family of RNA-binding proteins (9). Three other closely related neural proteins, called HelN1 [or HuB (10,11)], HuC (11) and HuD (12), are target antigens in paraneoplastic neuropathy [Hu syndrome (13)]. HuR is a 36-kDa polypeptide that contains three classical RNA-binding domains [RNA recognition motifs (RRMs)] (8): the first two have been implicated in ARE recognition, whereas the third has been suggested to bind the poly(A) tail of target mRNAs (14).…”

Section: Hur Binding To Cytoplasmic Mrna Is Perturbed By Heat Shockmentioning

confidence: 99%

HuR binding to cytoplasmic mRNA is perturbed by heat shock

Gallouzi

Brennan

Stenberg

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

298

280

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AU-rich elements (AREs) located in the 3′ untranslated region target the mRNAs encoding many protooncoproteins, cytokines, and lymphokines for rapid degradation. HuR, a ubiquitously expressed member of the embryonic lethal abnormal vision (ELAV) family of RNA-binding proteins, binds ARE sequences and selectively stabilizes ARE-containing reporter mRNAs when overexpressed in transiently transfected cells. HuR appears predominantly nucleoplasmic but has been shown to shuttle between the nucleus and cytoplasm via a novel shuttling sequence HNS. We report generation of a mouse monoclonal antibody 3A2 that both immunoblots and immunoprecipitates HuR protein; it recognizes an epitope located in the first of HuR's three RNA recognition motifs. This antibody was used to probe HuR interactions with mRNA before and after heat shock, a condition that has been reported to stabilize ARE-containing mRNAs. At 37°C, approximately one-third of the cytoplasmic HuR appears polysome associated, and in vivo UV crosslinking reveals that HuR interactions with poly(A) + RNA are predominantly cytoplasmic rather than nuclear. This comprises evidence that HuR directly interacts with mRNA in vivo . After heat shock, 12–15% of HuR accumulates in discrete foci in the cytoplasm, but surprisingly the majority of HuR crosslinks instead to nuclear poly(A) + RNA, whose levels are dramatically increased in the stressed cells. This behavior of HuR differs from that of another ARE-binding protein, hnRNP D, which has been implicated as an effector of mRNA decay rather than mRNA stabilization and of the general pre-RNA-binding protein hnRNP A1. We interpret these differences to mean that the temporal association of HuR with ARE-containing mRNAs is different from that of these other two proteins.

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Section: Hur Binding To Cytoplasmic Mrna Is Perturbed By Heat Shockmentioning

confidence: 99%

HuR binding to cytoplasmic mRNA is perturbed by heat shock

Gallouzi

Brennan

Stenberg

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

298

280

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“…Furthermore, a substantial body of evidence indicates that ELAV-like proteins exert a positive regulatory function on gene expression through posttranscriptional stabilization (10, 11) and possibly enhanced translation (12) of ARE-endowed mRNAs. One target of ELAV-like proteins is the mRNA of the GAP-43 gene (25), known to be a determinant of neural development, regeneration of neural connections, and synaptic plasticity in mature synapses (26). Whereas GAP-43 neuron-specific expression is controlled by elements in the promoter of this gene, its induction during neuritogenesis has been ascribed essentially to posttranscriptional events involving mRNA stabilization (27).…”

Section: Resultsmentioning

confidence: 99%

Posttranscriptional regulation of gene expression in learning by the neuronal ELAV-like mRNA-stabilizing proteins

Quattrone

Pascale

Nogués

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

130

125

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The view that memory is encoded by variations in the strength of synapses implies that long-term biochemical changes take place within subcellular microdomains of neurons. These changes are thought ultimately to be an effect of transcriptional regulation of specific genes. Localized changes, however, cannot be fully explained by a purely transcriptional control of gene expression. The neuron-specific ELAV-like HuB, HuC, and HuD RNA-binding proteins act posttranscriptionally by binding to adenine-and uridinerich elements (AREs) in the 3 untranslated region of a set of target mRNAs, and by increasing mRNA cytoplasmic stability and͞or rate of translation. Here we show that neuronal ELAV-like genes undergo a sustained up-regulation in hippocampal pyramidal cells only of mice and rats that have learned a spatial discrimination paradigm. This learning-specific increase of ELAV-like proteins was localized within cytoplasmic compartments of the somata and proximal dendrites and was associated with the cytoskeleton. This increase was also accompanied by enhanced expression of the GAP-43 gene, known to be regulated mainly posttranscriptionally and whose mRNA is demonstrated here to be an in vivo ELAV-like target. Antisense-mediated knockdown of HuC impaired spatial learning performance in mice and induced a concomitant downregulation of GAP-43 expression. Neuronal ELAV-like proteins could exert learning-induced posttranscriptional control of an array of target genes uniquely suited to subserve substrates of memory storage.

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“…Although this element contains several highly conserved U-rich stretches (A /GUUUG/ C), replacement of the middle U with A in three of the U-rich stretches did not change the half-life of the transcripts, suggesting that additional sequences in this region may contribute to its destabilizing effect. In fact, we recently have found a 25 nt U-rich region downstream of these motifs that contains a recognition site for a neuronal-specific elav-like RNA-binding protein [Chung et al (1997); also see below].…”

Section: Discussionmentioning

confidence: 99%

Post-Transcriptional Regulation of the GAP-43 Gene by Specific Sequences in the 3′ Untranslated Region of the mRNA

et al. 1997

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We have shown previously that GAP-43 gene expression during neuronal differentiation is controlled by selective changes in mRNA stability. This process was found to depend on highly conserved sequences in the 3Ј untranslated region (3Ј UTR) of the mRNA. To map the sequences in the GAP-43 3Ј UTR that mediate this post-transcriptional event, we generated specific 3Ј UTR deletion mutants and chimeras with the ␤-globin gene and measured their half-lives in transfected PC12 cells. Our results indicate that there are two distinct instability-conferring elements localized at the 5Ј and 3Ј ends of the GAP-43 3Ј UTR. Of these destabilizing elements, only the one at the 3Ј end is required for the stabilization of the mRNA in response to treatment with the phorbol ester TPA. This 3Ј UTR element consists of highly conserved uridine-rich sequences and contains specific recognition sites for two neural-specific GAP-43 mRNAbinding proteins. Analysis of the levels of mRNA and protein derived from various 3Ј UTR deletion mutants indicated that all mutants were translated effectively and that differences in gene expression in response to TPA were attributable to changes in GAP-43 mRNA stability. In addition, the phorbol ester was found to affect the binding of specific RNA-binding proteins to the 3Ј UTR of the GAP-43 mRNA. Given that, like the GAP-43 mRNA, its degradation machinery and the GAP-43 mRNAbinding proteins are expressed primarily in neural cells, we propose that these factors may be involved in the posttranscriptional regulation of GAP-43 gene expression during neuronal differentiation.

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The Elav-like Proteins Bind to a Conserved Regulatory Element in the 3′-Untranslated Region of GAP-43 mRNA

Cited by 141 publications

References 41 publications

HuR binding to cytoplasmic mRNA is perturbed by heat shock

HuR binding to cytoplasmic mRNA is perturbed by heat shock

Posttranscriptional regulation of gene expression in learning by the neuronal ELAV-like mRNA-stabilizing proteins

Post-Transcriptional Regulation of the GAP-43 Gene by Specific Sequences in the 3′ Untranslated Region of the mRNA

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