The cyclin-dependent kinase inhibitor p21 is regulated by RNA-binding protein PCBP4 via mRNA stability

ABSTRACT Expression of the cyclin-dependent kinase inhibitor p21 is highly induced by many stresses, including exposure to short-wavelength UV light (UVC), which increases p21 mRNA stability. Investigation into the mechanisms underlying this stabilization process revealed that proteins present in cytoplasmic lysates of human RKO colorectal carcinoma cells formed complexes with p21 mRNA that were inducible by treatment with UVC and other stress agents. The ubiquitous Elav-type RNA-binding protein HuR was identified within the p21 mRNA-protein complexes, as antibodies recognizing HuR supershifted these complexes and revealed HuR-immunoreactive proteins complexing with p21 mRNA on Western blots. Lowering of endogenous HuR levels through expression of antisense HuR decreased p21 RNA-protein complexes, greatly reduced the UVC inducibility and half-life of p21 mRNA, and prevented UVC-mediated induction of luciferase activity in p21 3′ untranslated region-containing reporter constructs. Our findings indicate that HuR plays a major role in regulating stress-induced p21 expression by enhancing p21 mRNA stability and that these effects are coupled to HuR's elevated presence in the cytoplasm.

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Faculty Opinions recommendation of Double-stranded RNA-binding protein regulates vascular endothelial growth factor mRNA stability, translation, and breast cancer angiogenesis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1104383.560401 ◽

2008 ◽

Author(s):

Lee Gehrke

Keyword(s):

Breast Cancer ◽

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Mrna Stability ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Vascular Endothelial ◽

Double Stranded Rna ◽

Endothelial Growth Factor

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Double-Stranded RNA-Binding Protein Regulates Vascular Endothelial Growth Factor mRNA Stability, Translation, and Breast Cancer Angiogenesis

Molecular and Cellular Biology ◽

10.1128/mcb.02078-06 ◽

2007 ◽

Vol 28 (2) ◽

pp. 772-783 ◽

Cited By ~ 77

Author(s):

Frank Vumbaca ◽

Kathryn N. Phoenix ◽

Daniel Rodriguez-Pinto ◽

David K. Han ◽

Kevin P. Claffey

Keyword(s):

Breast Cancer ◽

Mrna Stability ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Vascular Endothelial ◽

Vegf Mrna ◽

Double Stranded Rna ◽

Hypoxic Conditions ◽

Endothelial Growth Factor

ABSTRACT Vascular endothelial growth factor (VEGF) is a key angiogenic factor expressed under restricted nutrient and oxygen conditions in most solid tumors. The expression of VEGF under hypoxic conditions requires transcription through activated hypoxia-inducible factor 1 (HIF-1), increased mRNA stability, and facilitated translation. This study identified double-stranded RNA-binding protein 76/NF90 (DRBP76/NF90), a specific isoform of the DRBP family, as a VEGF mRNA-binding protein which plays a key role in VEGF mRNA stability and protein synthesis under hypoxia. The DRBP76/NF90 protein binds to a human VEGF 3′ untranslated mRNA stability element. RNA interference targeting the DRBP76/NF90 isoform limited hypoxia-inducible VEGF mRNA and protein expression with no change in HIF-1-dependent transcriptional activity. Stable repression of DRBP76/NF90 in MDA-MB-435 breast cancer cells demonstrated reduced polysome-associated VEGF mRNA levels under hypoxic conditions and reduced mRNA stability. Transient overexpression of the DRBP76/NF90 protein increased both VEGF mRNA and protein levels synthesized under normoxic and hypoxic conditions. Cells with stable repression of the DRBP76/NF90 isoform showed reduced tumorigenic and angiogenic potential in an orthotopic breast tumor model. These data demonstrate that the DRBP76/NF90 isoform facilitates VEGF expression by promoting VEGF mRNA loading onto polysomes and translation under hypoxic conditions, thus promoting breast cancer growth and angiogenesis in vivo.

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Integrative Regulatory Mapping Indicates that the RNA-Binding Protein HuR Couples Pre-mRNA Processing and mRNA Stability

Molecular Cell ◽

10.1016/j.molcel.2011.06.007 ◽

2011 ◽

Vol 43 (3) ◽

pp. 327-339 ◽

Cited By ~ 425

Author(s):

Neelanjan Mukherjee ◽

David L. Corcoran ◽

Jeffrey D. Nusbaum ◽

David W. Reid ◽

Stoyan Georgiev ◽

...

Keyword(s):

Mrna Stability ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Mrna Processing

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A Novel Role for the RNA–Binding Protein FXR1P in Myoblasts Cell-Cycle Progression by Modulating p21/Cdkn1a/Cip1/Waf1 mRNA Stability

PLoS Genetics ◽

10.1371/journal.pgen.1003367 ◽

2013 ◽

Vol 9 (3) ◽

pp. e1003367 ◽

Cited By ~ 46

Author(s):

Laetitia Davidovic ◽

Nelly Durand ◽

Olfa Khalfallah ◽

Ricardo Tabet ◽

Pascal Barbry ◽

...

Keyword(s):

Cell Cycle ◽

Mrna Stability ◽

Cell Cycle Progression ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Cycle Progression

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Nucleolar GTP-binding Protein-1 (NGP-1) Promotes G1to S Phase Transition by Activating Cyclin-dependent Kinase Inhibitor p21Cip1/Waf1

Journal of Biological Chemistry ◽

10.1074/jbc.m115.637280 ◽

2015 ◽

Vol 290 (35) ◽

pp. 21536-21552 ◽

Cited By ~ 8

Author(s):

Debduti Datta ◽

Kumaraswamy Anbarasu ◽

Suryaraja Rajabather ◽

Rangasamy Sneha Priya ◽

Pavitra Desai ◽

...

Keyword(s):

Phase Transition ◽

Kinase Inhibitor ◽

Binding Protein ◽

S Phase ◽

Cyclin Dependent Kinase ◽

Gtp Binding Protein ◽

Cyclin Dependent Kinase Inhibitor ◽

Gtp Binding

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Embryonic Lethal Abnormal Vision-like HuR-dependent mRNA Stability Regulates Post-transcriptional Expression of Cyclin-dependent Kinase Inhibitor p27Kip1

Journal of Biological Chemistry ◽

10.1074/jbc.m110.113365 ◽

2010 ◽

Vol 285 (20) ◽

pp. 15408-15419 ◽

Cited By ~ 20

Author(s):

Gudrun Ziegeler ◽

Jie Ming ◽

Jana C. Koseki ◽

Sema Sevinc ◽

Ting Chen ◽

...

Keyword(s):

Mrna Stability ◽

Kinase Inhibitor ◽

Cyclin Dependent Kinase ◽

Transcriptional Expression ◽

Cyclin Dependent Kinase Inhibitor ◽

Embryonic Lethal

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The RNA binding protein neuro‐oncological ventral antigen 1 (NOVA1) regulates IL-6 mRNA stability to enhance JAK2-STAT3 signaling in CRC

Surgical Oncology ◽

10.1016/j.suronc.2019.09.009 ◽

2019 ◽

Vol 31 ◽

pp. 67-74 ◽

Cited By ~ 2

Author(s):

Yong-gang Hong ◽

Guo-shu Xu ◽

Guan-yu Yu ◽

Ji-dian Zhou ◽

Qi-zhi Liu ◽

...

Keyword(s):

Mrna Stability ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Stat3 Signaling

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RPSAP52 lncRNA Inhibits p21Waf1/CIP Expression by Interacting With the RNA Binding Protein HuR

Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics ◽

10.3727/096504019x15761465603129 ◽

2020 ◽

Vol 28 (2) ◽

pp. 191-201

Author(s):

Daniela D’Angelo ◽

Claudio Arra ◽

Alfredo Fusco

Keyword(s):

Cell Cycle ◽

Pituitary Adenomas ◽

Noncoding Rna ◽

Rna Binding ◽

Kinase Inhibitor ◽

Cell Transformation ◽

Binding Protein ◽

Critical Role ◽

Rna Binding Protein ◽

Messenger Rnas

Long noncoding RNAs have been recently demonstrated to have an important role in fundamental biological processes, and their deregulated expression has been found in several human neoplasias. Our group has recently reported a drastic overexpression of the long noncoding RNA (lncRNA) RPSAP52 (ribosomal protein SA pseudogene 52) in pituitary adenomas. We have shown that this lncRNA increased cell proliferation by upregulating the expression of the chromatinic proteins HMGA1 and HMGA2, functioning as a competing endogenous RNA (ceRNA) through competitively binding to microRNA-15a (miR-15a), miR-15b, and miR-16. The aim of this work was to identify further mechanisms by which RPSAP52 overexpression could contribute to the development of pituitary adenomas. We investigated the involvement of RPSAP52 in the modulation of the expression of cell cycle-related genes, such as p21Waf1/CIP, whose deregulation plays a critical role in pituitary cell transformation. We report that RPSAP52, interacting with the RNA binding protein HuR (human antigen R), favors the delocalization of miR-15a, miR-15b, and miR-16 on the cyclin-dependent kinase inhibitor p21Waf1/CIP1 that, accordingly, results in downregulation in pituitary adenomas. A RNA immunoprecipitation sequencing (RIPseq) analysis performed on cells overexpressing RPSAP52 identified 40 messenger RNAs (mRNAs) enriched in Argonaute 2 (AGO2) immunoprecipitated samples. Among them, we focused on GAS8 (growth arrest-specific protein 8) gene. Consistently, GAS8 expression was downregulated in all the analyzed pituitary adenomas with respect to normal pituitary and in RPSAP52-overepressing cells, supporting the role of RPSAP52 in addressing genes involved in growth inhibition and cell cycle arrest to miRNA-induced degradation. This study unveils another RPSAP52-mediated molecular mechanism in pituitary tumorigenesis.

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