| 1997 |
KSRP was purified and cloned as a component of a multiprotein complex binding to an intronic splicing enhancer downstream of the neuron-specific c-src N1 exon; antibodies to KSRP inhibited N1 exon splicing in vitro, and this inhibition was rescued by addition of purified KSRP; KSRP contains four KH RNA-binding domains and induces assembly of five other proteins including hnRNP F onto the splicing enhancer. |
Protein purification, cDNA cloning, in vitro splicing assay with antibody inhibition and rescue by purified protein, co-purification of multiprotein complex |
Genes & development |
High |
9136930
|
| 2004 |
KSRP is an essential factor for ARE-directed mRNA decay; specific KH domains directly mediate RNA binding, mRNA decay, and physical interactions with the exosome and poly(A) ribonuclease (PARN); the ability of KH motifs to promote mRNA decay correlates with their ability to bind the ARE and associate with RNA-degrading enzymes. |
In vitro mRNA decay assays, domain mutagenesis, co-immunoprecipitation of KSRP with exosome and PARN, ARE-binding assays |
Molecular cell |
High |
15175153
|
| 2005 |
p38α and p38β MAP kinase phosphorylates KSRP during muscle differentiation; p38-phosphorylated KSRP displays compromised binding to ARE-containing myogenic transcripts and fails to promote their rapid decay, although it retains ability to interact with mRNA degradation machinery; this stabilizes critical myogenic mRNAs and is required for proper myoblast-to-myocyte conversion. |
In vitro kinase assay (p38 phosphorylation of KSRP), RNA-binding assays with phosphorylated vs. unphosphorylated KSRP, KSRP overexpression in myoblasts with transcriptional and post-transcriptional readouts |
Molecular cell |
High |
16364914
|
| 2005 |
KSRP binds the 3'-UTR ARE of human iNOS mRNA and promotes its decay by recruiting the exosome; KSRP competes with the stabilizing factor HuR for the same ARE binding site; KSRP also co-immunoprecipitates with tristetraprolin (TTP); a non-binding KSRP mutant fails to reduce iNOS expression, confirming the requirement for ARE binding. |
Immunodepletion in vitro degradation assays, overexpression of wild-type and non-binding mutant KSRP, co-immunoprecipitation with exosome and TTP, competition binding assay with HuR |
Nucleic acids research |
High |
16126846
|
| 2006 |
AKT phosphorylates KSRP at a unique serine residue; this induces KSRP association with 14-3-3 proteins and prevents KSRP interaction with the exosome, impairing its ability to promote rapid mRNA decay; this mechanism stabilizes beta-catenin mRNA downstream of PI3K-AKT and Wnt signaling. |
In vitro AKT kinase assay, co-immunoprecipitation of KSRP with 14-3-3 and exosome under PI3K-AKT activation/inhibition, mRNA half-life measurements, mutagenesis of phosphorylation site |
PLoS biology |
High |
17177604
|
| 2006 |
Tethering KSRP to mRNAs via fusion with sequence-specific RNA-binding proteins elicits mRNA decay in a manner dependent on known mRNA decay enzymes; this demonstrates that KSRP functions by recruiting mRNA decay machinery to target transcripts. |
Tethering assay using Rev-KSRP fusion protein targeting HIV-1 RRE; mRNA stability measurements; viral replication assays |
Molecular and cellular biology |
Medium |
16648466
|
| 2007 |
The KH3 and KH4 domains of KSRP behave as independent RNA-binding modules recognizing different regions of AU-rich RNA targets; KH4 binds AREs with lower affinity than KH3; mRNA binding and mRNA degradation activities of KSRP are closely associated with a conserved structural element unique to KH4. |
NMR structure determination of KH3 and KH4, RNA-binding affinity measurements (ITC/NMR), mutagenesis of KH4 structural element |
Structure |
High |
17437720
|
| 2007 |
KSRP knockdown by siRNA stabilizes IL-8 mRNA and impairs its rapid deadenylation; KSRP interacts with both domains of the bipartite IL-8 ARE; IL-1 treatment impairs the interaction of KSRP with the IL-8 ARE in a p38-dependent but MK2-independent manner; microarray pulldown identified ~100 mRNAs controlled by KSRP. |
siRNA knockdown, mRNA half-life and deadenylation assays, RNA pulldown with KSRP followed by microarray, co-immunoprecipitation, pharmacological inhibition of p38/MK2 |
Molecular and cellular biology |
High |
17908789
|
| 2008 |
Among the four KH domains of KSRP, KH3 binds RNA with the highest affinity and specifically recognizes G-rich sequences; the other KH domains display different sequence preferences explaining the broad target range; KSRP shows strong negative selectivity for sequences with multiple adjacent cytosines. |
Scaffold-independent RNA-binding analysis of all four KH domains using NMR and ITC; systematic sequence selectivity assays |
Nucleic acids research |
High |
18684992
|
| 2009 |
KSRP is a component of both Drosha and Dicer complexes; KSRP binds with high affinity to the terminal loop of target miRNA precursors and promotes their maturation; this function is required for specific changes in target mRNA expression affecting proliferation, apoptosis, and differentiation. |
Co-immunoprecipitation of KSRP with Drosha and Dicer complexes, RNA immunoprecipitation, in vitro processing assays, KSRP knockdown with miRNA profiling, NMR binding studies |
Nature |
High |
19458619
|
| 2009 |
AKT-mediated phosphorylation of KSRP's KH1 domain causes KH1 unfolding (KH1 is structurally atypical and unstable), creating a binding site for 14-3-3ζ; 14-3-3ζ binding discriminates phosphorylated from unphosphorylated KH1 and drives nuclear localization of KSRP, sequestering it from cytoplasmic mRNA decay activity. |
NMR structure of KH1 (phosphorylated and unphosphorylated), 14-3-3ζ binding assays, localization studies, mRNA decay assays in cells with phosphomimetic/phosphodead mutants |
Nature structural & molecular biology |
High |
19198587
|
| 2009 |
KSRP co-immunoprecipitates with PTH mRNA, the exosome, and the endoribonuclease PMR1; KSRP and exosome knockdown by siRNA prevents PMR1-mediated cleavage of PTH mRNA, establishing that PTH mRNA decay by PMR1 is dependent on both KSRP and the exosome. |
Co-immunoprecipitation of KSRP, PMR1, and exosome; siRNA knockdown of KSRP and exosome subunits; in vitro RNA cleavage assays |
BMC cell biology |
Medium |
19775426
|
| 2010 |
hnRNP A1 binds the terminal loop of pri-let-7a-1 and inhibits Drosha processing; hnRNP A1 binding to let-7a interferes with KSRP binding to the same terminal loop, establishing antagonistic roles for hnRNP A1 (negative regulator) and KSRP (positive regulator) in let-7a biogenesis. |
RNA-protein binding competition assays, in vitro Drosha processing assays, cell-based depletion/overexpression of hnRNP A1 and KSRP with miRNA quantification |
Nature structural & molecular biology |
High |
20639884
|
| 2010 |
KSRP interacts with Dishevelled (Dvl) proteins; KSRP negatively regulates Wnt/beta-catenin signaling by promoting post-transcriptional destabilization of CTNNB1 (beta-catenin) mRNA; identified by affinity purification coupled to ion-trap mass spectrometry. |
Affinity purification followed by mass spectrometry identification of KSRP as Dvl-binding protein; mRNA stability assays for beta-catenin mRNA; co-immunoprecipitation |
Journal of cell science |
Medium |
20332102
|
| 2010 |
The inter-domain linker between KH2 and KH3 orients the two central KH domains of KSRP with their RNA-binding surfaces aligned, creating a two-domain unit; this interdomain arrangement is functionally important for interaction with KSRP's RNA targets. |
NMR structural analysis of interdomain linker and relative domain orientation; RNA binding assays comparing wild-type and linker-mutated constructs |
Nucleic acids research |
High |
20385598
|
| 2011 |
KSRP is required for ARE-mediated decay of Ifna4 and Ifnb mRNAs; KSRP knockout mouse embryonic fibroblasts produce higher levels of type I IFN mRNAs following viral infection due to decreased mRNA decay; increased IFN expression renders Ksrp-/- cells refractory to HSV-1 and VSV infection. |
Ksrp knockout mouse MEFs, viral infection, mRNA half-life measurements, RNA immunoprecipitation, antiviral protection assays |
Molecular and cellular biology |
High |
21690298
|
| 2011 |
KSRP participates in ARE-mediated translational silencing in addition to mRNA decay; KSRP depletion increases polysome association of IL-6 and IL-1α mRNAs; silencing depends on the 3'-UTR ARE of IL-6 mRNA; IL-1 reverses KSRP-dependent translational silencing. |
Polysome profiling after siRNA-mediated KSRP depletion, luciferase reporter assays with IL-6 3'UTR, KSRP-RNA interaction mapping |
The Journal of biological chemistry |
Medium |
21795706
|
| 2011 |
PI3K/AKT activation regulates two distinct KSRP functions in opposite directions during skeletal myogenesis: it inhibits KSRP's ability to promote decay of myogenin mRNA while simultaneously activating its ability to favor maturation of myogenic miRNAs; this dynamic switch is required for the myogenic program as demonstrated in C2C12 cells and Ksrp knockout mice. |
C2C12 myoblast differentiation, Ksrp-/- mouse analysis, PI3K/AKT inhibition/activation, miRNA processing assays, mRNA stability measurements |
Cell death and differentiation |
High |
21886180
|
| 2012 |
KSRP KH3 domain recognizes a G-rich sequence in the pre-let-7 terminal loop via a noncanonical KH-RNA interaction mode; the KH3-RNA complex structure reveals how this noncanonical binding governs KSRP selectivity for let-7 precursors; this selectivity is crucial for the functional interaction between KSRP and let-7 biogenesis. |
NMR structure of KH3 bound to pre-let-7 terminal loop G-rich sequence, mutagenesis of KH3 and RNA, functional processing assays |
Nature structural & molecular biology |
High |
23142982
|
| 2012 |
HuR promotes myogenesis by associating with the decay factor KSRP and ribonucleases PARN and the exosome to destabilize nucleophosmin (NPM) mRNA; the C terminus of HuR mediates formation of the HuR-KSRP complex; this HuR-KSRP-PARN-exosome complex reduces NPM mRNA levels required for muscle cell commitment. |
Co-immunoprecipitation of HuR-KSRP complex, mRNA stability assays, domain deletion analysis of HuR, siRNA knockdown of HuR, overexpression studies |
Nature communications |
Medium |
24969639
|
| 2012 |
Phosphorylated R-SMAD proteins (transducers of BMP2 signaling) associate with phosphorylated KSRP and block its interaction with primary myogenic miRNAs (myomiRs), preventing KSRP-dependent myomiR maturation; silencing SMAD4, SMAD5, and SMAD9 rescues KSRP function; this SMAD-mediated KSRP blockade is required for BMP2-induced osteoblastic differentiation. |
Co-immunoprecipitation of phospho-SMAD with phospho-KSRP, RNA immunoprecipitation, siRNA knockdown of SMADs, miRNA processing assays, cell differentiation readouts |
Cell reports |
Medium |
23177623
|
| 2013 |
KSRP destabilizes GAP-43 mRNA by binding its ARE, a process requiring its KH4 domain; KSRP competes with the stabilizing factor HuD for binding to GAP-43 ARE sequences; KSRP overexpression hinders axonal outgrowth in primary hippocampal neurons, while KSRP knockdown increases GAP-43 mRNA and dramatically increases axonal length in both cultured neurons and Ksrp+/- and Ksrp-/- embryos. |
RNA-binding assays (KH domain deletion mutants), competition assays with HuD, overexpression/knockdown in primary hippocampal neurons, Ksrp knockout mouse neurons, axonal length measurements |
PloS one |
High |
24244461
|
| 2013 |
KSRP promotes decay of Per2 mRNA via AU-rich elements in Per2's 3' UTR, and KSRP ablation increases Per2 expression causing a phase delay in circadian clock gene oscillations and reduced expression of de novo lipogenic genes, leading to reduced hepatic triglyceride accumulation. |
Ksrp-/- mouse liver analysis, heterologous mRNA reporter assays with Per2 3'UTR, mRNA half-life measurements in primary hepatocytes, circadian gene profiling |
Journal of lipid research |
Medium |
25514904
|
| 2013 |
p38 activation increases utrophin A mRNA stability through AU-rich elements by reducing the functional availability of KSRP; KSRP directly binds AREs in the utrophin A 3'UTR; electroporation-mediated knockdown of KSRP increases utrophin A in wild-type and mdx mouse muscles. |
In vitro and in vivo KSRP-RNA binding studies, mRNA half-life assays, electroporation knockdown in mouse muscle, p38 activation with heparin in mdx mice |
Human molecular genetics |
Medium |
23575223
|
| 2013 |
The RNA helicase DDX1 interacts with KSRP and regulates its subcellular localization; DDX1 knockdown elevates cytoplasmic levels of KSRP and facilitates ARE-mediated mRNA decay; KSRP associates with DDX1 or 14-3-3 proteins in a mutually exclusive manner, indicating competing interactions that regulate KSRP's cytoplasmic availability. |
Affinity purification and mass spectrometry identification of DDX1, co-immunoprecipitation, DDX1 siRNA knockdown, subcellular fractionation, mRNA decay assays |
PloS one |
Medium |
24023901
|
| 2014 |
KSRP directly binds the H19 lncRNA in the cytoplasm of undifferentiated C2C12 cells, and this interaction facilitates KSRP-mediated destabilization of labile transcripts including myogenin mRNA; AKT activation induces KSRP dismissal from H19; upon dismissal, KSRP is repurposed to promote maturation of myogenic miRNAs; H19 thus acts as a molecular scaffold that optimizes an AKT-regulated post-transcriptional switch. |
RNA immunoprecipitation, in vitro RNA-protein binding assays, AKT activation/inhibition, mRNA stability assays, miRNA processing assays in C2C12 cells |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
25385579
|
| 2014 |
KSRP ablation causes a reduction in body adiposity; in Ksrp-/- inguinal WAT, expression of miR-150 is decreased due to impaired primary miR-150 processing; miR-150 directly targets PRDM16 and PPARGC1A; forced miR-150 expression attenuates elevated brown fat gene expression caused by KSRP deletion, establishing a pathway KSRP → miR-150 → PRDM16/PPARGC1A controlling brown-like transformation of white fat. |
Ksrp-/- mouse analysis, miRNA processing assays, miRNA mimic rescue experiments, miR-150 target validation by luciferase assay |
Diabetes |
Medium |
24722250
|
| 2014 |
In Ksrp-/- epididymal WAT, miR-145 expression is decreased due to impaired primary miR-145 processing; miR-145 directly targets and represses FOXO1 and CGI58, activators of lipolysis; forced miR-145 expression attenuates enhanced lipolysis in KSRP-deficient adipose tissue, establishing a KSRP → miR-145 → FOXO1/CGI58 pathway controlling adipose lipolysis. |
Ksrp-/- mouse WAT analysis, miRNA processing assays, miR-145 target validation by luciferase assay, miR-145 mimic rescue in primary adipocytes |
Molecular and cellular biology |
Medium |
24732799
|
| 2016 |
KHSRP is ubiquitinated at Lys109, Lys121, and Lys122 by the Kelch domain of KLHL12 interacting with KHSRP's C-terminal domain; mutations at these sites abolish KHSRP's inhibitory effect on enterovirus IRES-driven translation; ubiquitination of KHSRP (a negative ITAF) thus controls IRES-dependent translation by limiting competition against positive ITAFs. |
In vivo and in vitro SUMOylation/ubiquitination assays, site-directed mutagenesis of ubiquitination sites, Ni2+-NTA pulldown, co-immunoprecipitation of KLHL12-KHSRP, IRES-luciferase reporter assays |
Nucleic acids research |
High |
27899653
|
| 2016 |
miR-27b-3p-mediated silencing of KHSRP is required for TGF-β-induced EMT in mammary gland cells; KHSRP controls levels of mature miR-192-5p (targeting EMT factors) and promotes epithelial-type alternative splicing of pre-mRNAs including Cd44 and Fgfr2; KHSRP and hnRNPA1 cooperate in a ribonucleoprotein complex to promote epithelial exon usage. |
miR-27b-3p overexpression/knockdown, KHSRP knockdown/overexpression, genome-wide RNA-seq and miRNA-seq, co-immunoprecipitation of KHSRP-hnRNPA1 complex, splicing assays |
Cell reports |
Medium |
27396342
|
| 2017 |
KHSRP is modified by SUMO1 at the major site K87; this modification inhibits KHSRP's interaction with pri-miRNA/Drosha-DGCR8 complex and promotes KHSRP translocation from nucleus to cytoplasm; SUMOylation consequently impairs processing of pre-miRNAs from pri-miRNAs with G-rich terminal loops (TL-G-Rich miRNAs including let-7 family), leading to tumorigenesis. |
In vivo and in vitro SUMOylation assays, site-directed mutagenesis (K87), nuclear/cytosol fractionation, immunofluorescence, RIP, high-throughput miRNA sequencing, xenograft tumor models |
Molecular cancer |
High |
29020972
|
| 2017 |
KHSRP associates with the regulatory domain of RIG-I to maintain the receptor in an inactive state and attenuate its sensing of viral RNA; KHSRP knockdown enhances RIG-I antiviral signaling and reduces viral replication both in vitro and in vivo. |
RNAi screens integrated with interactome data, co-immunoprecipitation of KHSRP with RIG-I regulatory domain, RIG-I signaling assays upon KHSRP knockdown, in vitro and in vivo viral infection models |
Nature microbiology |
Medium |
28248290
|
| 2002 |
Xenopus KSRP homolog VgRBP71 binds to mRNAs localized to either vegetal or animal hemispheres in oocytes but does not accumulate at the vegetal cortex with the mRNA; VgRBP71 interacts with Prrp (another Vg1 localization element-binding protein) in a manner that does not require Vg1 mRNA. |
Xenopus oocyte expression library screen, in vivo RNA-protein binding assays, co-immunoprecipitation of VgRBP71-Prrp interaction, localization studies |
Development |
Medium |
12421702
|
| 2002 |
KSRP/FBP2 is cleaved by caspase-3 and caspase-7 at two sites: after Asp102 and after Asp183; the second cleavage site is only recognized in protein produced in cells (not in vitro translated protein), indicating that cellular context beyond primary sequence affects caspase recognition. |
In vitro caspase cleavage of KSRP with caspase-3 and -7, comparison of cellular vs. in vitro translated KSRP, site identification by mass spectrometry |
Protein and peptide letters |
Medium |
12553859
|
| 2022 |
Khsrp mRNA localizes into peripheral nerve axons and is locally translated following axon injury; axonal KSRP levels rapidly increase after sciatic nerve injury and remain elevated; KHSRP binds to mRNAs with 3'UTR AU-rich elements (Gap43, Snap25, Fubp1) and targets them to the cytoplasmic exosome for degradation; Ksrp-/- mice show increased axonal levels of these target mRNAs and accelerated nerve regeneration in vivo. |
Targeted proteomics of axoplasm, Ksrp-/- mouse nerve injury model, local translation assay with axon ligation, RNA immunoprecipitation, mRNA stability measurements in axons, in vivo nerve regeneration assay |
Nucleic acids research |
High |
35556128
|
| 2022 |
KHSRP regulates monocytic differentiation through both transcriptional and post-transcriptional mechanisms; KHSRP occupies specific genomic sites at promoter and enhancer regions to regulate hematopoietic gene transcription, and binds to pre-mRNA intronic regions to modulate alternative splicing; KHSRP has co-regulatory effects at both transcriptional and post-transcriptional levels on MOGOH and ADARB1. |
ChIP-seq, RNA-seq, CLIP-seq in CD34+ cells during monocytic differentiation, functional knockdown studies |
Blood science |
Medium |
36518592
|
| 2024 |
KHSRP directly interacts with SF3B1 (splicing factor 3b subunit 1) and enhances SF3B1 binding to intronic branch sites, thereby promoting pre-mRNA splicing; KHSRP knockdown causes intron retention and exacerbates liver injury in acute liver failure models. |
Co-immunoprecipitation of KHSRP-SF3B1 interaction, RNA-seq analysis of splicing defects in KHSRP-knockdown cells and murine ALF models, splicing inhibitor experiments in vivo, proteomic and transcriptomic analyses |
Cell death & disease |
Medium |
39187547
|
| 2024 |
KHSRP functions as an m6A-binding protein; KHSRP recognizes m6A modifications and stabilizes FAK pathway mRNAs including MET, ITGAV, and ITGB1 in an m6A-dependent manner, leading to activation of downstream FAK signaling and promotion of pancreatic ductal adenocarcinoma progression. |
m6A-RIP, RNA immunoprecipitation, KHSRP knockdown/overexpression with mRNA stability assays, FAK pathway activity assays, in vivo xenograft models, PROTAC-mediated KHSRP degradation |
Cancer research |
Medium |
39120596
|
| 2024 |
SIRT7 is a deacetylase of KHSRP; androgen stimulation impairs SIRT7 interaction with KHSRP, leading to increased KHSRP acetylation; acetylated KHSRP has impaired mRNA decay activity for DDR-related mRNAs, promoting prostate cancer tumorigenesis. |
Co-immunoprecipitation of SIRT7-KHSRP interaction, acetylation assays under androgen stimulation, RNA-seq and GSEA for DDR gene changes, xenograft tumor growth with acetylation-site mutants |
Molecular oncology |
Medium |
38501452
|
| 2015 |
KHSRP and hnRNPA1 form a ribonucleoprotein complex and cooperate to promote epithelial-type exon usage during splicing; combined silencing of both prevents RESV-dependent inclusion of epithelial-type exons in Cd44 pre-mRNA; hnRNP A1 also tethers KSRP to an exonic splicing silencer (ESS16) and KSRP binding to ESS16 requires hnRNP A1. |
Co-immunoprecipitation of KHSRP-hnRNPA1 complex, combined siRNA knockdown, splicing assays, 2D gel and mass spectrometry identification in erythroleukemia cells |
American journal of cancer research / Biochimica et biophysica acta |
Medium |
26101706 28088441
|