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Cited by 110RNA Research and SplicingCholesterol and Lipid MetabolismRNA Interference and Gene Delivery

The DOI matches its Crossref record; 31 of the 31 references it cites were found in a registry or an index; 17 of the 31 sources its claims cite were checked for support.

ZFP36L1 and ZFP36L2 control LDLR mRNA stability via the ERK–RSK pathway

Open access
Adachi, Shungo, Homoto, Masae, Tanaka, Rikou, Hioki, Yusaku, Murakami, Hiroshi, Suga, Hiroaki, Matsumoto, Masaki, Nakayama, Keiichi I., Hatta, Tomohisa, Iemura, Shun-ichiro, Natsume, Tohru·2014·Nucleic Acids Research
DOI 10.1093/nar/gku652PMC PMC4150769
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Abstract

Low-density lipoprotein receptor (LDLR) mRNA is unstable, but is stabilized upon extracellular signal-regulated kinase (ERK) activation, possibly through the binding of certain proteins to the LDLR mRNA 3′-untranslated region (UTR), although the detailed mechanism underlying this stability control is unclear. Here, using a proteomic approach, we show that proteins ZFP36L1 and ZFP36L2 specifically bind to the 3′-UTR of LDLR mRNA and recruit the CCR4-NOT-deadenylase complex, resulting in mRNA destabilization. We also show that the C-terminal regions of ZFP36L1 and ZFP36L2 are directly phosphorylated by p90 ribosomal S6 kinase, a kinase downstream of ERK, resulting in dissociation of the CCR4-NOT-deadenylase complex and stabilization of LDLR mRNA. We further demonstrate that targeted disruption of the interaction between LDLR mRNA and ZFP36L1 and ZFP36L2 using antisense oligonucleotides results in upregulation of LDLR mRNA and protein. These results indicate that ZFP36L1 and ZFP36L2 regulate LDLR protein levels downstream of ERK. Our results also show the usefulness of our method for identifying critical regulators of specific RNAs and the potency of antisense oligonucleotide-based therapeutics.

Overview

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Written from the paper’s full text.

TL;DR

ZFP36L1 and ZFP36L2 bind the LDLR mRNA 3′-UTR ARE1 region and recruit the CCR4-NOT deadenylase complex to destabilize LDLR mRNA; ERK–RSK signaling phosphorylates their C-termini, dissociating CCR4-NOT and stabilizing LDLR mRNA. LNA antisense oligonucleotide Oligo-L1 disrupts this interaction, increasing LDLR mRNA/protein and LDL uptake in Hep3B cells.

Key findings

  • A proteomic screen using seven Flag-tagged bait RNAs identified ZFP36L1 and ZFP36L2 as specific binders of the LDLR mRNA 3′-UTR (ARE1–3); about 400 peptides from ∼150 proteins were identified, with ∼25% common to all baits, and ZFP36L1/L2 appeared in all four LDLR MS analyses. Endogenous ZFP36L1 co-immunoprecipitated LDLR, PLK3, and VEGFA mRNAs but not β-actin mRNA.
  • ZFP36L1 and ZFP36L2 predominantly bound the ARE1 UAUUUAUU sequence. Double siRNA knockdown of ZFP36L1 and ZFP36L2 in HeLa cells increased LDLR mRNA and protein and stabilized LDLR mRNA; PMA-mediated LDLR mRNA stabilization was lost after knockdown. LNA Oligo-L1 blocked the LDLR–ZFP36L1/L2 interaction, increased LDLR mRNA and protein in HeLa and Hep3B cells, and increased DiI-LDL uptake in Hep3B cells; control oligos L2–L6 did not.
  • ERK-dependent phosphorylation was mapped to C-terminal serine-334 of ZFP36L1 and serine-493/495 of ZFP36L2 by iTRAQ MS; PMA increased these phosphorylations and U0126 reversed them. PMA inhibited ZFP36L1–CNOT7 interaction, reversed by U0126 or RSK inhibitors BI-D1870/SL0101; recombinant RSK1 directly phosphorylated GST-ZFP36L1 at serine-334 and reduced CNOT1/CNOT7 binding.
  • ZFP36L1-SASA or ZFP36L1-ΔC mutants prevented PMA-mediated stabilization of an RFP-LDLR-3′-UTR reporter. PLK3 and VEGFA mRNAs were also stabilized by PMA, whereas cMYC mRNA was not.

Methods

  • In vitro RNA-binding protein purification: seven Flag-peptide-tagged bait RNAs, including LDLR 3′-UTR fragments, stable RNAs, and c-Myc, were incubated with 293T lysates, immunoprecipitated, and analyzed by nanoscale LC/MS/MS; interactions were validated by western blot and qPCR co-immunoprecipitation.
  • Cell-based assays in HEK293T, HeLa, and Hep3B cells: siRNA knockdown (20 nM), LNA-modified antisense oligonucleotides (80 nM), actinomycin D chase with or without PMA, qPCR normalized to β-actin or Luc2, mRNA half-lives fitted by exponential decay, western blot, and DiI-LDL uptake in Hep3B cells.
  • Phosphorylation analysis: iTRAQ-based quantitative MS of Flag-ZFP36L1/L2 from mock-, PMA-, or PMA+U0126-treated 293T cells; in vitro kinase assays with recombinant RSK1/ERK1 and GST-ZFP36L1; Student’s t-test with P < 0.002, n = 3 for qPCR groups.

Limitations

  • The paper states that the mechanisms and meaning of PMA-enhanced ZFP36L1 binding to LDLR mRNA remain unknown, and whether ZFP36 family proteins are commonly regulated by these pathways or differ is an open question.
  • The paper states that UAUUUAUU is necessary but not sufficient for ZFP36L1/L2 interaction and destabilization, and that further investigation is required to predict target mRNAs in silico; off-target effects of antisense oligonucleotides are important to consider for therapeutic applications.
  • Evident from the text: no in vivo animal or human data are presented, and delivery of Oligo-L1 to the liver is not tested; the in vitro RBP purification approach may not fully capture in vivo RNA–protein interactions, although the authors argue it has advantages.

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Integrity signals

Registry

Crossref · 10.1093/nar/gku652

Cites

31 of 31 references verified0 citation errors · 0 not checkable

Statistics

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Figures

Figure 1. Identification of bait-specific RBPs. (A) Flow chart summarizing the experiment. (B) Confirmation of our results by western blot analysis. 293T cells were lysed in lysis buffer and the cleared lysates were subjected to IP with anti-Flag antibody in combination with the indicated bait RNAs. Co-immunoprecipitated RNA and proteins were eluted using the Flag-peptide and were then subjected to western blot analysis using the indicated antibodies. Five percent of the initial amount of cleared 293T lysate was loaded as total lysate. An IP experiment without bait RNA was performed as a control. (C) Confirmation of the endogenous interaction between ZFP36L1 and LDLR mRNA. 293T cells were lysed in lysis buffer and the cleared lysates were subjected to IP with control or anti-ZFP36L1 antibody. Total RNA and co-immunoprecipitated RNA were extracted, and quantitative reverse-transcription (RT)-PCR (qPCR) was performed using primers specific to LDLR, PLK3, VEGFA and β-actin mRNAs. Results are shown as% of input. The data are representative of at least three independent experiments. Error bars show standard deviation of the mean.
Figure 1. Identification of bait-specific RBPs. (A) Flow chart summarizing the experiment. (B) Confirmation of our results by western blot analysis. 293T cells were lysed in lysis buffer and the cleared lysates were subjected to IP with anti-Flag antibody in combination with the indicated bait RNAs. Co-immunoprecipitated RNA and proteins were eluted using the Flag-peptide and were then subjected to western blot analysis using the indicated antibodies. Five percent of the initial amount of cleared 293T lysate was loaded as total lysate. An IP experiment without bait RNA was performed as a control. (C) Confirmation of the endogenous interaction between ZFP36L1 and LDLR mRNA. 293T cells were lysed in lysis buffer and the cleared lysates were subjected to IP with control or anti-ZFP36L1 antibody. Total RNA and co-immunoprecipitated RNA were extracted, and quantitative reverse-transcription (RT)-PCR (qPCR) was performed using primers specific to LDLR, PLK3, VEGFA and β-actin mRNAs. Results are shown as% of input. The data are representative of at least three independent experiments. Error bars show standard deviation of the mean.CC BY · PubMed Central
Figure 2. Domain analysis of the 3′-UTR of LDLR mRNA. (A) Schematic representation of the 3′-UTR of LDLR mRNA. Black bars indicate the regions used for ActD chase and IP experiments. AREs 1–3 are indicated by red boxes and the UCAU repeat is indicated by the white box. Designed oligonucleotides are indicated by blue lines. Conservation around the LDLR ARE1 region is shown in the blue graph, created using Vertebrate Multiz Alignment & PhastCons Conservation (28 species), as extracted from the UCSC Human Genome Browser (http://genome.ucsc.edu/). The consensus ZFP36L1- and ZFP36L2-binding sequence is colored in red. Sequences of the designed oligonucleotides, L1 and L2, are indicated. (B) RFP-tagged LDLR 3′-UTR constructs were expressed in 293T cells, and cells were then subjected to ActD chase experiments. Twenty-four hours after transfection, 293T cells were treated with ActD and PMA as indicated (PMA treatment commenced 10 min after ActD treatment). qPCR was performed using RFP and Luc2-specific primers. Results were normalized to the levels of Luc2 mRNA. The data are representative of at least three independent experiments. mRNA half-lives were calculated by fitting an exponential decay curve to all time points. Figures in the graph indicate the mean value of the mRNA decay half-lives. Error bars show standard deviation of the mean. P-values were calculated using Student's t-test. *P < 0.002; n = 3 for each group. (C) 293T cells were transfected with Myc-tagged ZFP36L1 and ZFP36L2. Cells were then lysed with lysis buffer and the cleared lysates were subjected to IP with the indicated LDLR 3′-UTR bait RNAs. Immunoprecipitated proteins were subjected to western blot analysis with the anti-Myc antibody. (D) 293T cells were transfected with Myc-tagged ZFP36L1 and ZFP36L2. Cells were then lysed with lysis buffer and the cleared lysates were subjected to IP with the indicated LDLR 3′-UTR bait RNAs and the indicated oligos. Immunoprecipitated proteins were subjected to western blot analysis with the anti-Myc antibody. These data are representative of at least three independent experiments.
Figure 2. Domain analysis of the 3′-UTR of LDLR mRNA. (A) Schematic representation of the 3′-UTR of LDLR mRNA. Black bars indicate the regions used for ActD chase and IP experiments. AREs 1–3 are indicated by red boxes and the UCAU repeat is indicated by the white box. Designed oligonucleotides are indicated by blue lines. Conservation around the LDLR ARE1 region is shown in the blue graph, created using Vertebrate Multiz Alignment & PhastCons Conservation (28 species), as extracted from the UCSC Human Genome Browser (http://genome.ucsc.edu/). The consensus ZFP36L1- and ZFP36L2-binding sequence is colored in red. Sequences of the designed oligonucleotides, L1 and L2, are indicated. (B) RFP-tagged LDLR 3′-UTR constructs were expressed in 293T cells, and cells were then subjected to ActD chase experiments. Twenty-four hours after transfection, 293T cells were treated with ActD and PMA as indicated (PMA treatment commenced 10 min after ActD treatment). qPCR was performed using RFP and Luc2-specific primers. Results were normalized to the levels of Luc2 mRNA. The data are representative of at least three independent experiments. mRNA half-lives were calculated by fitting an exponential decay curve to all time points. Figures in the graph indicate the mean value of the mRNA decay half-lives. Error bars show standard deviation of the mean. P-values were calculated using Student's t-test. *P < 0.002; n = 3 for each group. (C) 293T cells were transfected with Myc-tagged ZFP36L1 and ZFP36L2. Cells were then lysed with lysis buffer and the cleared lysates were subjected to IP with the indicated LDLR 3′-UTR bait RNAs. Immunoprecipitated proteins were subjected to western blot analysis with the anti-Myc antibody. (D) 293T cells were transfected with Myc-tagged ZFP36L1 and ZFP36L2. Cells were then lysed with lysis buffer and the cleared lysates were subjected to IP with the indicated LDLR 3′-UTR bait RNAs and the indicated oligos. Immunoprecipitated proteins were subjected to western blot analysis with the anti-Myc antibody. These data are representative of at least three independent experiments.CC BY · PubMed Central
Figure 3. LDLR mRNA is destabilized by ZFP36L1 and ZFP36L2. (A) HeLa cells were transfected with ZFP36L1 and ZFP36L2 siRNAs. Forty-eight hours after transfection, cells were harvested, total RNA was extracted and quantitative RT-PCR (qPCR) was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to β-actin mRNA levels. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test.*P < 0.002; n = 3 for each group. (B) Forty-eight hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (C) Forty-eight hours after transfection, cells were treated with ActD and chased for the indicated time with or without PMA (PMA treatment commenced 10 min after ActD treatment). Total RNA was extracted and qPCR was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to the levels of β-actin mRNA. Error bars show standard deviation of the mean. (D) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and total RNA was extracted. Quantitative RT-PCR (qPCR) was performed using primers specific for LDLR and β-actin. Results were normalized to β-actin mRNA levels. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test. *P < 0.002; n = 3 for each group. (E) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (F) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were treated with ActD and chased for the indicated times. Total RNA was extracted and qPCR was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to the levels of β-actin mRNA. Error bars show standard deviation of the mean. (G) Hep3B cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (H) Hep3B cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were treated with DiI-LDL for 1 h. The cells were then lysed in RIPA buffer and the ratio of DiI-LDL fluorescence/protein concentration was measured. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test. *P < 0.002; n = 3 for each group. The data are representative of at least three independent experiments. The number below the figure indicates the number of times we replicated the experiment. Data from one of the independent experiments are shown in Supplementary Figure S4A–H.
Figure 3. LDLR mRNA is destabilized by ZFP36L1 and ZFP36L2. (A) HeLa cells were transfected with ZFP36L1 and ZFP36L2 siRNAs. Forty-eight hours after transfection, cells were harvested, total RNA was extracted and quantitative RT-PCR (qPCR) was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to β-actin mRNA levels. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test.*P < 0.002; n = 3 for each group. (B) Forty-eight hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (C) Forty-eight hours after transfection, cells were treated with ActD and chased for the indicated time with or without PMA (PMA treatment commenced 10 min after ActD treatment). Total RNA was extracted and qPCR was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to the levels of β-actin mRNA. Error bars show standard deviation of the mean. (D) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and total RNA was extracted. Quantitative RT-PCR (qPCR) was performed using primers specific for LDLR and β-actin. Results were normalized to β-actin mRNA levels. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test. *P < 0.002; n = 3 for each group. (E) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (F) HeLa cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were treated with ActD and chased for the indicated times. Total RNA was extracted and qPCR was performed using primers specific to LDLR mRNA and β-actin mRNA. Results were normalized to the levels of β-actin mRNA. Error bars show standard deviation of the mean. (G) Hep3B cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were harvested and the lysates were subjected to western blot analysis using the indicated antibodies. (H) Hep3B cells were transfected with the indicated oligos. Twenty-four hours after transfection, cells were treated with DiI-LDL for 1 h. The cells were then lysed in RIPA buffer and the ratio of DiI-LDL fluorescence/protein concentration was measured. Error bars show standard deviation of the mean. P-values against control were calculated using Student's t-test. *P < 0.002; n = 3 for each group. The data are representative of at least three independent experiments. The number below the figure indicates the number of times we replicated the experiment. Data from one of the independent experiments are shown in Supplementary Figure S4A–H.CC BY · PubMed Central
Figure 4. ERK pathway phosphorylates and inhibits ZFP36L2. (A) 293T cells were transfected with Flag-tagged ZFP36L1. Twenty-four hours after transfection, cells were treated with PMA (for 30 min) and U0126 (for 45 min), as indicated. Cell lysates were subjected to western blot analysis using anti-Flag antibody. (B) 293T cells were transfected with Flag-tagged ZFP36L1. Cells were treated with or without PMA (for 30 min) and the lysates were subjected to IP. Immunopurified Flag-ZFP36L1 was treated with or without bacterial acid phosphatase (BAP) and samples were subjected to western blot analysis using anti-Flag antibody. (C) Alignment of C-terminal amino acid sequences in the human ZFP36 family. The C-terminal end of each protein is shown as (*). Identical residues are shown by (|) and similar residues are shown by (+). The serine residues identified as being phosphorylated downstream of ERK are shown in red. (D) 293T cells were transfected with Myc-tagged ZFP36L1 or ZFP36L2 and then treated with or without PMA (for 30 min). Cell lysates were subjected to IP using the Flag-antibody and the indicated bait RNAs. Co-immunoprecipitated proteins were subjected to western blot analysis using anti-MYC (9E10) antibody. (E) 293T cells were transfected with Flag-tagged ZFP36L1, then treated with PMA (for 30 min) and U0126 (for 45 min) as indicated, and lysates were subjected to IP. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. (F) Schematic representation of the constructs used in this experiment. (G) 293T cells were transfected with the indicated constructs. Twenty-four hours after transfection, cells were treated with or without PMA (for 30 min), and lysates were subjected to IP. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. (H) HeLa cells were transfected with RFP-LDLR 3′-UTR (ARE1) or Luc2-β-Actin-UTR along with the indicated ZFP36L1 constructs. Twenty-four hours after transfection, cells were treated with ActD and PMA (PMA treatment commenced 15 min after ActD treatment), as indicated. Two hours after ActD treatment, cells were harvested and total RNA was extracted. Quantitative RT-PCR (qPCR) was performed using RFP and Luc2-specific primers. Results were normalized to the levels of Luc2 mRNA. The data are representative of at least three independent experiments.
Figure 4. ERK pathway phosphorylates and inhibits ZFP36L2. (A) 293T cells were transfected with Flag-tagged ZFP36L1. Twenty-four hours after transfection, cells were treated with PMA (for 30 min) and U0126 (for 45 min), as indicated. Cell lysates were subjected to western blot analysis using anti-Flag antibody. (B) 293T cells were transfected with Flag-tagged ZFP36L1. Cells were treated with or without PMA (for 30 min) and the lysates were subjected to IP. Immunopurified Flag-ZFP36L1 was treated with or without bacterial acid phosphatase (BAP) and samples were subjected to western blot analysis using anti-Flag antibody. (C) Alignment of C-terminal amino acid sequences in the human ZFP36 family. The C-terminal end of each protein is shown as (*). Identical residues are shown by (|) and similar residues are shown by (+). The serine residues identified as being phosphorylated downstream of ERK are shown in red. (D) 293T cells were transfected with Myc-tagged ZFP36L1 or ZFP36L2 and then treated with or without PMA (for 30 min). Cell lysates were subjected to IP using the Flag-antibody and the indicated bait RNAs. Co-immunoprecipitated proteins were subjected to western blot analysis using anti-MYC (9E10) antibody. (E) 293T cells were transfected with Flag-tagged ZFP36L1, then treated with PMA (for 30 min) and U0126 (for 45 min) as indicated, and lysates were subjected to IP. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. (F) Schematic representation of the constructs used in this experiment. (G) 293T cells were transfected with the indicated constructs. Twenty-four hours after transfection, cells were treated with or without PMA (for 30 min), and lysates were subjected to IP. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. (H) HeLa cells were transfected with RFP-LDLR 3′-UTR (ARE1) or Luc2-β-Actin-UTR along with the indicated ZFP36L1 constructs. Twenty-four hours after transfection, cells were treated with ActD and PMA (PMA treatment commenced 15 min after ActD treatment), as indicated. Two hours after ActD treatment, cells were harvested and total RNA was extracted. Quantitative RT-PCR (qPCR) was performed using RFP and Luc2-specific primers. Results were normalized to the levels of Luc2 mRNA. The data are representative of at least three independent experiments.CC BY · PubMed Central
Figure 5. RSK1 directly phosphorylates the C-terminus of ZFP36L1. (A) 293T cells were transfected with Flag-tagged ZFP36L1. Twenty-four hours after transfection, cells were treated with PMA (for 30 min) and BI-D1870 or SL0101 (for 45 min) as indicated, and lysates were subjected to IP with the anti-Flag antibody. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. Five percent of the initial amount of cleared 293T lysate was loaded as total lysate. (B) GST-ZFP36L1 protein was incubated with or without active recombinant RSK1 under phosphorylation conditions for 30 min. Phosphorylation of GST-ZFP36L1 C-terminal peptide was analyzed by MS. The MS/MS spectrum used to identify the m/z 681.8604-peak as the LPIFSRLSISDD peptide fragment of ZFP36L1 is shown. (C) MS/MS spectrum used to identify the m/z 721.8421-peak as the serine-8 phosphorylated LPIFSRLSISDD peptide fragment of ZFP36L1. (D) GST-ZFP36L1 protein was incubated with the indicated recombinant kinase under phosphorylation conditions for 30 min, washed, then added to 293T cell lysate and then subjected to pulldown with glutathione-sepharose. Eluted proteins were subjected to western blot analysis using the indicated antibodies and subjected to Coomassie Brilliant Blue (CBB) staining. The asterisk shows a nonspecific band. Five percent of the initial amount of cleared 293T cell lysate was loaded as total lysate. These data are representative of at least three independent experiments.
Figure 5. RSK1 directly phosphorylates the C-terminus of ZFP36L1. (A) 293T cells were transfected with Flag-tagged ZFP36L1. Twenty-four hours after transfection, cells were treated with PMA (for 30 min) and BI-D1870 or SL0101 (for 45 min) as indicated, and lysates were subjected to IP with the anti-Flag antibody. Co-immunoprecipitated proteins were subjected to western blot analysis using the indicated antibodies. Five percent of the initial amount of cleared 293T lysate was loaded as total lysate. (B) GST-ZFP36L1 protein was incubated with or without active recombinant RSK1 under phosphorylation conditions for 30 min. Phosphorylation of GST-ZFP36L1 C-terminal peptide was analyzed by MS. The MS/MS spectrum used to identify the m/z 681.8604-peak as the LPIFSRLSISDD peptide fragment of ZFP36L1 is shown. (C) MS/MS spectrum used to identify the m/z 721.8421-peak as the serine-8 phosphorylated LPIFSRLSISDD peptide fragment of ZFP36L1. (D) GST-ZFP36L1 protein was incubated with the indicated recombinant kinase under phosphorylation conditions for 30 min, washed, then added to 293T cell lysate and then subjected to pulldown with glutathione-sepharose. Eluted proteins were subjected to western blot analysis using the indicated antibodies and subjected to Coomassie Brilliant Blue (CBB) staining. The asterisk shows a nonspecific band. Five percent of the initial amount of cleared 293T cell lysate was loaded as total lysate. These data are representative of at least three independent experiments.CC BY · PubMed Central

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41 of 41

  • The paper relies on this without support.

    RSK is downstream of ERK.

    “p90 ribosomal S6 kinase, a kinase downstream of ERK”

  • The paper relies on this without support.

    ARE1 and ARE2 of the LDLR mRNA 3′-UTR are comprised of the UAUUUAUU sequence.

    “ARE1 and ARE2 are comprised of the UAUUUAUU sequence.”

  • The paper relies on this without support.

    The UAUUUAUU sequence is the consensus binding sequence for ZFP36L1 and ZFP36L2.

    “ZFP36L1 and ZFP36L2 are known as proteins that bind to a certain type of ARE that contains the sequence UAUUUAUU, causing destabilization of target mRNAs.”

  • The paper relies on this without support.

    LDLR in the liver binds LDL and thereby lowers blood LDL levels.

    “In the liver, the LDLR binds to LDL, thereby lowering blood LDL levels.”

  • The paper relies on this without support.

    cMYC mRNA does not bind to ZFP36L1 or ZFP36L2.

    “whereas cMYC mRNA does not bind to ZFP36L1 or ZFP36L2.”
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Metadata

Authors
Adachi, S., Homoto, M., Tanaka, R., Hioki, Y., Murakami, H., Suga, H., Matsumoto, M., Nakayama, K. I., Hatta, T., Iemura, S.-I., & Natsume, T.
Affiliations
  • National Institute of Advanced Industrial Science and Technology (JP)
  • The University of Tokyo (JP)
  • Kyushu University (JP)
Corresponding author
Natsume, Tohru
ORCID
Venue
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Published
2014-08-08
Volume
42
Issue
15
Pages
10037-10049
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
DOI
10.1093/nar/gku652
PMC
PMC4150769
OpenAlex
W2019768782
Cited by
109
References
30
Citations by year
2026: 9 · 2025: 10 · 2024: 20 · 2023: 9 · 2022: 7 · 2021: 16 · 2020: 9 · 2019: 8 · 2018: 6 · 2017: 3 · 2016: 3 · 2015: 8 · 2014: 1
FWCI
1.65
Type
journal article
Language
en
Open access
Licence
Locations
  • Nucleic Acids Research (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institute of Advanced Industrial Science and Technology
  • Japan Society for the Promotion of Science (26640080, 24115001)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
ZFP36L1, ZFP36L2, LDL receptor, mRNA stability, ERK signaling, p90 ribosomal S6 kinase, CCR4-NOT complex, mRNA deadenylation, 3' untranslated region, protein phosphorylation, antisense oligonucleotides, lipid metabolism
MeSH terms
Cell Line, HeLa Cells, Humans, Phosphorylation, Receptors, LDL, RNA, Messenger, Transcription Factors, 3' Untranslated Regions, RNA Stability, MAP Kinase Signaling System, Ribosomal Protein S6 Kinases, 90-kDa, Extracellular Signal-Regulated MAP Kinases, Butyrate Response Factor 1, HEK293 Cells

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