Embryonic Lethal Abnormal Vision-Like RNA-Binding Proteins Regulate Neurite Outgrowth and Tau Expression in PC12 Cells

Aranda-Abreu, Gonzálo Emiliano; Behar, Leah; Chung, Sangmi; Furneaux, Henry; Ginzburg, Irith

doi:10.1523/jneurosci.19-16-06907.1999

Cited by 138 publications

(144 citation statements)

References 66 publications

Supporting

Mentioning

138

Contrasting

Unclassified

Order By: Relevance

“…Besides GAP-43, HuD is known to bind the 3Ј UTRs of other mRNAs expressed in neurons, such as those for c-fos, tau, N-myc, and p21 waf1 (Chung et al, 1997;Joseph et al, 1998;Aranda-Abreu et al, 1999;Lazarova et al, 1999). Therefore, it is likely that the expression of other genes may be affected by the presence of HuD antisense or sense transcripts.…”

Section: Discussionmentioning

confidence: 99%

The RNA-binding Protein HuD Is Required for GAP-43 mRNA Stability, GAP-43 Gene Expression, and PKC-dependent Neurite Outgrowth in PC12 Cells

Mobarak¹,

Anderson²,

Morin³

et al. 2000

MBoC

Self Cite

126

146

View full text Add to dashboard Cite

The RNA-binding protein HuD binds to a regulatory element in the 3Ј untranslated region (3Ј UTR) of the GAP-43 mRNA. To investigate the functional significance of this interaction, we generated PC12 cell lines in which HuD levels were controlled by transfection with either antisense (pDuH) or sense (pcHuD) constructs. pDuH-transfected cells contained reduced amounts of GAP-43 protein and mRNA, and these levels remained low even after nerve growth factor (NGF) stimulation, a treatment that is normally associated with protein kinase C (PKC)-dependent stabilization of the GAP-43 mRNA and neuronal differentiation. Analysis of GAP-43 mRNA stability demonstrated that the mRNA had a shorter half-life in these cells. In agreement with their deficient GAP-43 expression, pDuH cells failed to grow neurites in the presence of NGF or phorbol esters. These cells, however, exhibited normal neurite outgrowth when exposed to dibutyryl-cAMP, an agent that induces outgrowth independently from GAP-43. We observed opposite effects in pcHuD-transfected cells. The GAP-43 mRNA was stabilized in these cells, leading to an increase in the levels of the GAP-43 mRNA and protein. pcHuD cells were also found to grow short spontaneous neurites, a process that required the presence of GAP-43. In conclusion, our results suggest that HuD plays a critical role in PKC-mediated neurite outgrowth in PC12 cells and that this protein does so primarily by promoting the stabilization of the GAP-43 mRNA. INTRODUCTIONIn addition to transcriptional factors, RNA-binding proteins play a critical role in the developmental control of gene expression. Among these is ELAV (embryonic lethal abnormal vision), an RNA-binding protein identified in Drosophila, where the gene is required for normal development and maintenance of the nervous system (Campos et al., 1985;Robinow et al., 1988). In higher vertebrates and mammals, four members of the ELAV-like family have been identified. These are also referred to as Hu proteins, namely HuR (also known as HuA), HuB (Hel-N1), HuC, and HuD, because these are targets of anti-Hu antibodies present in the sera of patients with paraneoplastic encephalomyelitis (Dalmau et al., 1992). HuR is ubiquitously expressed (Ma et al., 1996), while HuB, HuC, and HuD are expressed uniquely in the nervous system. Recent studies indicate that overexpression of neural ELAV-like proteins is sufficient to induce neuronal differentiation in vitro and in vivo (Wakamatsu and Weston, 1997; Akamatsu et al., 1999;Antic et al., 1999;Kasashima et al., 1999). While the exact function and targets of ELAV/Hu proteins remain to be fully elucidated, it seems likely that this family of RNA-binding proteins controls neuronal differentiation by selectively modulating the expression of neural-specific, growth-associated genes.The growth-associated protein GAP-43 is expressed in neurons primarily during the initial establishment and regeneration of neural connections (Skene, 1989;Benowitz and Routtenberg, 1997 Eggen et al., 1995;Chiaramello et al., 1996;Kinney et ...

show abstract

Section: Discussionmentioning

confidence: 99%

The RNA-binding Protein HuD Is Required for GAP-43 mRNA Stability, GAP-43 Gene Expression, and PKC-dependent Neurite Outgrowth in PC12 Cells

Mobarak¹,

Anderson²,

Morin³

et al. 2000

MBoC

Self Cite

126

146

View full text Add to dashboard Cite

show abstract

“…This result opened the intriguing possibility that dozens of ELAV͞Hu targeted mRNAs containing AREs could be stabilized and͞or translationally activated as a group in response to ELAV͞Hu protein. mRNAs shown to be affected following overexpression of Hu proteins include glucose trans-porter 1 (12), NF-M (13), GAP43 (15), VEGF (16), c-fos (17)(18)(19), c-myc (unpublished results), TNF-␣ (19), GM-CSF (19), and tau (20). With the exception of NF-M mRNA (13), the binding of these mRNA targets to ELAV͞Hu proteins has only been demonstrated when using in vitro methods.…”

Section: Elav͞hu Rna Recognition Motif (Rrm) Proteins Associate With Amentioning

confidence: 97%

Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome

Keene

2001

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

216

240

View full text Add to dashboard Cite

“…This indicates that the Hu proteins share similar target preferences. Thus, AChE joins a growing family of transcripts that contain varying types of AREs that are stabilized by the Hu family of proteins in several distinct tissues (40,41,49,60,61).…”

Section: Fig 6 Hur Overexpression Increases Ache Mrna Stabilitymentioning

confidence: 99%

The RNA-binding Protein HuR Binds to Acetylcholinesterase Transcripts andRegulates Their Expression in Differentiating Skeletal MuscleCells

Deschênes‐Furry

Bélanger

Mwanjewe

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

During myogenic differentiation, acetylcholinesterase (AChE) transcript levels are known to increase dramatically. Although this increase can be attributed in part to increased transcriptional activity, posttranscriptional mechanisms have also been implicated in the high levels of AChE mRNA in myotubes. In this study, we observed that transfection of a luciferase reporter construct containing the full-length AChE 3-untranslated region (UTR) resulted in significantly higher (5-fold) luciferase activity in differentiated myotubes versus myoblasts. RNA-electrophoretic mobility shift assays (REMSAs) performed with a full-length AChE 3-UTR probe and the AU-rich element revealed that the intensity of RNA-binding protein complexes increased as myogenic differentiation proceeded. Using several complementary approaches including supershift REMSA, mRNA-binding protein pull-down assays, and immunoprecipitation followed by reverse transcription-PCR, we found that the mRNA-stabilizing protein HuR interacts directly with AChE transcripts. Stable overexpression of HuR in C2C12 cells increased the expression of endogenous AChE transcripts as well as that of the luciferase reporter construct containing the AChE 3-UTR. In vitro stability assays performed with protein extracts from these cells versus controls resulted in a slower rate of AChE mRNA decay. The down-regulation of HuR expression mediated through small interfering RNA further confirmed the role of HuR in the regulation of AChE mRNA levels. Taken together, these studies demonstrate that HuR interacts with the AChE 3-UTR to regulate posttranscriptionally the expression of AChE mRNA during myogenic differentiation.Neurotransmission at cholinergic synapses of the central and peripheral nervous systems is promptly terminated by the catalytic activity of the enzyme acetylcholinesterase (AChE)

show abstract

Embryonic Lethal Abnormal Vision-Like RNA-Binding Proteins Regulate Neurite Outgrowth and Tau Expression in PC12 Cells

Cited by 138 publications

References 66 publications

The RNA-binding Protein HuD Is Required for GAP-43 mRNA Stability, GAP-43 Gene Expression, and PKC-dependent Neurite Outgrowth in PC12 Cells

The RNA-binding Protein HuD Is Required for GAP-43 mRNA Stability, GAP-43 Gene Expression, and PKC-dependent Neurite Outgrowth in PC12 Cells

Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome

The RNA-binding Protein HuR Binds to Acetylcholinesterase Transcripts andRegulates Their Expression in Differentiating Skeletal MuscleCells

Contact Info

Product

Resources

About