"We also show that phospho-KH1 interacts directly and specifically with 14-3-3 in vitro and phosphorylation of Ser193 causes a 14-3-3-dependent alteration of the nuclear/cytoplasmic distribution of the protein."
"To confirm that this model is applicable to our data and to show that the 14-3-3-KH1 interaction is indeed mediated by a short unfolded and phosphorylated amino acid stretch we have used a 10-fold excess of a phospho-peptide that spans the KH114-3-3 target sequence (GLPERSVS p LTGAPES) to compete out 14-3-3 protein."
"The experiments described above show that KH1 interacts with 14-3-3 only when the domain is phosphorylated and unfolded and that, as we previously proposed xref , this interaction is mediated by a short target sequence within the domain."
"The data demonstrate interaction of isolated PCBP2KH1 and KH3 domains to four distinct target sites within the 5'-nontranslated region of the CVB3 genomic RNA."
"In this model, the extensive interactions of PCBP2's KH1 domain with d10c would clearly be critical for its function and would account for the strict requirement for this domain in initiation on Type 1 IRESs ( xref , xref )."
"When we re-evaluated the interaction of PCBP2 with MAVS, we found that MAVS specifically interacted with the KH1 domain of PCBP2, which is the same domain that bound EPRS ( xref and Supplementary Fig. 7a,b )."
"We also show that phospho-KH1 interacts directly and specifically with 14-3-3 in vitro and phosphorylation of Ser193 causes a 14-3-3-dependent alteration of the nuclear/cytoplasmic distribution of the protein."
"To confirm that this model is applicable to our data and to show that the 14-3-3-KH1 interaction is indeed mediated by a short unfolded and phosphorylated amino acid stretch we have used a 10-fold excess of a phospho-peptide that spans the KH114-3-3 target sequence (GLPERSVS p LTGAPES) to compete out 14-3-3 protein."
"The experiments described above show that KH1 interacts with 14-3-3 only when the domain is phosphorylated and unfolded and that, as we previously proposed xref , this interaction is mediated by a short target sequence within the domain."
"The data demonstrate interaction of isolated PCBP2KH1 and KH3 domains to four distinct target sites within the 5'-nontranslated region of the CVB3 genomic RNA."
"In this model, the extensive interactions of PCBP2's KH1 domain with d10c would clearly be critical for its function and would account for the strict requirement for this domain in initiation on Type 1 IRESs ( xref , xref )."
"When we re-evaluated the interaction of PCBP2 with MAVS, we found that MAVS specifically interacted with the KH1 domain of PCBP2, which is the same domain that bound EPRS ( xref and Supplementary Fig. 7a,b )."
"In contrast to KH1, resonances from the S1 and KH2 regions were not affected, suggesting that AR2 interacts exclusively with the KH1 domain (Figure xref , xref )."
"Addition of alphaCTD to the SKK : AR2 complex reversed the chemical shift changes induced in the KH1 signals on AR2 addition (XREF_FIG), confirming that alphaCTD promotes NusA binding to RNA by displacing AR2 from SKK."
"Our results revealed that break of PCBP1KH1 domain apparently mitigated p27 protein expression and the reporter activity, but not the KH2 and KH3 mutations, indicating that PCBP1 binds to p27 mRNA mainly through its KH1 domain, rather than KH2 and KH3 domains, which is consistent to that PCBP1KH1 domain interacts with stem-loops I and IV of EV71 5’UTR, facilitating viral replication [ xref ]."
"Thus, we proposed that KH1 domain of PCBP1 specifically binds to stem-loop I and IV of EV71 5′UTR, which may play an important biological function in viral replication."
"Our results revealed that break of PCBP1KH1 domain apparently mitigated p27 protein expression and the reporter activity, but not the KH2 and KH3 mutations, indicating that PCBP1 binds to p27 mRNA mainly through its KH1 domain, rather than KH2 and KH3 domains, which is consistent to that PCBP1KH1 domain interacts with stem-loops I and IV of EV71 5’UTR, facilitating viral replication [ xref ]."
"KH1 binding to KSRP AU-rich target sequences is significantly weaker than the binding of the other domains (at least 10-fold weaker than KH3), while the differences between KH2, KH3 and KH4 are only 2- to 3-fold ( xref )."
"KH1 binding to KSRP AU rich target sequences is significantly weaker than the binding of the other domains (at least 10-fold weaker than KH3), while the differences between KH2, KH3 and KH4 are only 2- to 3-fold."
"To further examine whether the interaction of KSRP with the exosome is necessary for RNA decay, the decay of ARE tnf was examined in the KSRP depleted S100 supplemented with KSRP and either GST-KH 3 or GST-KH 1."
"In contrast to KH-1 (valence = 0, and 1.2), which only triggered moderate apoptosis, KH-1 with higher valences of 5.4, 9.4 and 16.6 significantly enhanced apoptosis, highlighting the importance of multivalence."
"Our results revealed that break of PCBP1KH1 domain apparently mitigated p27 protein expression and the reporter activity, but not the KH2 and KH3 mutations, indicating that PCBP1 binds to p27 mRNA mainly through its KH1 domain, rather than KH2 and KH3 domains, which is consistent to that PCBP1KH1 domain interacts with stem-loops I and IV of EV71 5’UTR, facilitating viral replication [ xref ]."
"Thus, we proposed that KH1 domain of PCBP1 specifically binds to stem-loop I and IV of EV71 5′UTR, which may play an important biological function in viral replication."
"Our results revealed that break of PCBP1KH1 domain apparently mitigated p27 protein expression and the reporter activity, but not the KH2 and KH3 mutations, indicating that PCBP1 binds to p27 mRNA mainly through its KH1 domain, rather than KH2 and KH3 domains, which is consistent to that PCBP1KH1 domain interacts with stem-loops I and IV of EV71 5’UTR, facilitating viral replication [ xref ]."
"KH1 binding to KSRP AU-rich target sequences is significantly weaker than the binding of the other domains (at least 10-fold weaker than KH3), while the differences between KH2, KH3 and KH4 are only 2- to 3-fold ( xref )."
"KH1 binding to KSRP AU rich target sequences is significantly weaker than the binding of the other domains (at least 10-fold weaker than KH3), while the differences between KH2, KH3 and KH4 are only 2- to 3-fold."
"Unfolding of KH1 does not impair the ability of KSRP to bind the mRNA, since the KH3 and KH4 domains of KSRP mediate high affinity RNA binding to the ARE in TNF-alpha."
"In contrast to KH1, resonances from the S1 and KH2 regions were not affected, suggesting that AR2 interacts exclusively with the KH1 domain (Figure xref , xref )."
"Two independent phosphorylation events are involved in KHSRP-mediated mRNA degradation: p38 MAPK phosphorylated C-terminal Thr692 regulated the decay of ARE-containing myogenic mRNAs, while AKT phosphorylated KH1 domain reduced the degradation rate of mRNAs [51,52] ."
"Our model for PRRSV −2/−1 PRF is that PCBPs interact with the PRRSV mRNA through KH1 and KH3, with each domain contacting one of the two C-patches (CCCA and CUCC) within the C-rich region."
"Furthermore, KH1 acts as a dominant negative mutant to inhibit translation from a poliovirus reporter gene in both Xenopus laevis oocytes and HeLa cell in vitro translation extracts."
"Although the inhibition of mTOR2 induced by GDC-0980 alone was not as significant as PI3K or mTOR1, further addition of KH-1 markedly decreased mTOR2 activity."
"Our model for PRRSV −2/−1 PRF is that PCBPs interact with the PRRSV mRNA through KH1 and KH3, with each domain contacting one of the two C-patches (CCCA and CUCC) within the C-rich region."
"In addition, GDC-0980 and KH-1 also worked together to down-regulate NF-kappaB and Bcl-2 which are broadly associated with oncogenesis through their ability to facilitate cell progression."
"For closely related CVB3, individual KH1 and KH3 domains of PCBP bind to IRES stem loop IV, KH1 interacts with subdomain IV/C RNA, whereas KH3 interacts with subdomain IV/B."
"Full-length CAGE, but not KH1 deletion construct, conferred resistance to trastuzumab in Malme3M cells, prevented caspase-3 activity from increasing in response to trastuzumab in Malme3M cells and prevented cleavage of PARP in response to trastuzumab in Malme3M cells."
"For closely related CVB3, individual KH1 and KH3 domains of PCBP bind to IRES stem loop IV, KH1 interacts with subdomain IV/C RNA, whereas KH3 interacts with subdomain IV/B."
"This analysis showed that ct Dim2 utilizes its KH-like domain (KH1) to interact with ct Utp1, which is reminiscent of how Dim2–KH1 binds to Nob1 xref ."
"The KH1 variant could bind MITF RNA, however its binding profile does not follow the usual saturation binding curve as exhibited by the WT CRD-BP, and hence its K d value can not be determined (XREF_FIG)."
"For example, Krr1 binds via its KH1 (KH-like) domain to Kri1, a nucleolar assembly factor associated with snR30 xref , whereas the KH1 domain of Dim2 provides a binding site for the endonuclease Nob1 xref ."
"Next, we have added FUSE DNA to the FIR RRM1-RRM2-FBP Nbox KH1 and KH4 complex described above and recorded again a fingerprint NMR 15 N- 1 H correlation NMR spectrum."
"In this assay, the KH1 and KH2 mutants increased pause dwell times similarly to wild-type NusA, supporting the view that interactions of the KH domains with the nascent RNA or with RNAP are not important for the pause enhancement by NusA (XREF_FIG; XREF_TABLE)."
"We propose that KH1 mediates transit of LmxGT1 from the flagellar pocket into the flagellar membrane via interaction with the proximal portion of the flagellar axoneme."
"As these drugs have different effects on inhibition of cell cycle with G2/M phase arrest by DTX [XREF_BIBR], S phase arrest by GEM [XREF_BIBR], and G1/G0 arrest by GDC-0980 [XREF_BIBR], we further investigated the impact of KH-1 on drug modulated cell cycle."
"In contrast, GDC-0980 apparently negated the calcium influx induced by KH-1 (XREF_FIG), which might be explained by PI3K dependent extracellular calcium influx [XREF_BIBR] and potential inhibition of calcium influx by PI3K inhibitors such as GDC-0980."
"This analysis showed that ct Dim2 utilizes its KH-like domain (KH1) to interact with ct Utp1, which is reminiscent of how Dim2–KH1 binds to Nob1 xref ."
"Our results also lend support to earlier mutational data which showed that RNA binding was abolished by mutating the first glycine in the conserved GXXG motifs of KH1 (G253D) and KH2 (G319D)."
"Full-length CAGE, but not KH1 deletion construct, conferred resistance to trastuzumab in Malme3M cells, prevented caspase-3 activity from increasing in response to trastuzumab in Malme3M cells and prevented cleavage of PARP in response to trastuzumab in Malme3M cells."
"The KH1 variant could bind MITF RNA, however its binding profile does not follow the usual saturation binding curve as exhibited by the WT CRD-BP, and hence its K d value can not be determined (XREF_FIG)."
"When we re-evaluated the interaction of PCBP2 with MAVS, we found that MAVS specifically interacted with the KH1 domain of PCBP2, which is the same domain that bound EPRS ( xref and Supplementary Fig. 7a,b )."
"We propose that KH1 mediates transit of LmxGT1 from the flagellar pocket into the flagellar membrane via interaction with the proximal portion of the flagellar axoneme."
"For example, Krr1 binds via its KH1 (KH-like) domain to Kri1, a nucleolar assembly factor associated with snR30 xref , whereas the KH1 domain of Dim2 provides a binding site for the endonuclease Nob1 xref ."
"Next, we have added FUSE DNA to the FIR RRM1-RRM2-FBP Nbox KH1 and KH4 complex described above and recorded again a fingerprint NMR 15 N- 1 H correlation NMR spectrum."
"Consistently, binding of all three KH domains was almost completely abolished by M7, M8 and M11 substitutions whereas binding of KH1 and KH2 was moderately reduced by M9 and M10 substitutions and binding of KH3 was unaffected."
"An in vitro binding assay revealed that PCBP2 KH1 specifically interacted with the GST and L1 regions of EPRS (amino acids 1–196) but not with the GST-like domain alone (amino acids 1–168) ( xref and Supplementary Fig. 6a,b ), which further supported the finding that EPRS L1 was essential for interaction with PCBP2."
"16 These intense research efforts on KH-1 and its derivatives have been driven by the apparently strong immune responses induced by KH-1 as compared to monomeric Le y and Le x antigens."
"The deletion analysis showing in the xref also raised a possibility of intramolecular interactions within SHI1 because KH2 inhibits the interactions of KH1 and KH3 with FRY2/CPL1."
"Full-length CAGE, but not KH1 deletion construct, increased the expression of pEGFR Y845 and HER2, and induced interaction of EGFR with CAGE and HER2."
"The activity of extracellular enzymes cellulase, amylase, and protease, and the amount of EPS produced by mutant strain KH1 were not significantly different from the wild type strain Xc1 (data not shown)."
"Full-length CAGE, but not KH1 deletion construct, increased the expression of pEGFR Y845 and HER2, and induced interaction of EGFR with CAGE and HER2."
"Interestingly, while KH-1 alone had little impact on cell cycle regulation, it exerted drug dependent effect on the alteration of cell cycle when combined with different drugs (XREF_FIG) : KH-1 enhanced DTX induced G2/M arrest, GEM induced S phase arrest and GDC-0980-induced G1/G0 arrest."
"In addition, GDC-0980 and KH-1 also worked together to down-regulate NF-kappaB and Bcl-2 which are broadly associated with oncogenesis through their ability to facilitate cell progression."
"Since 2-NAc-NAN was not demethylated by strain KH1 (XREF_TABLE), metabolites 208a and 208b are proposed to result from demethylation of DNAN followed by two successive acetylations of the primary amine group."
"An in vitro binding assay revealed that PCBP2 KH1 specifically interacted with the GST and L1 regions of EPRS (amino acids 1–196) but not with the GST-like domain alone (amino acids 1–168) ( xref and Supplementary Fig. 6a,b ), which further supported the finding that EPRS L1 was essential for interaction with PCBP2."
"The deletion analysis showing in the xref also raised a possibility of intramolecular interactions within SHI1 because KH2 inhibits the interactions of KH1 and KH3 with FRY2/CPL1."
"The results can be summarized as follows : the KH1 and KH3 domains bind the BAT element (XREF_SUPPLEMENTARY, right upper panel), whereas KH1 and KH2 domains bind eEF1A1 (XREF_SUPPLEMENTARY, right bottom panel)."
"Our results revealed that break of PCBP1KH1 domain apparently mitigated p27 protein expression and the reporter activity, but not the KH2 and KH3 mutations, indicating that PCBP1 binds to p27 mRNA mainly through its KH1 domain, rather than KH2 and KH3 domains, which is consistent to that PCBP1KH1 domain interacts with stem-loops I and IV of EV71 5’UTR, facilitating viral replication [ xref ]."
"Although both the KH1 and the KH3 domains of PCBP1 bound to BolA2 in an Fe- and GSH dependent manner, only KH3 could support the formation of [2Fe-2S] clusters on the BolA2 and Glrx3 complex in cells."
"The results can be summarized as follows : the KH1 and KH3 domains bind the BAT element (XREF_SUPPLEMENTARY, right upper panel), whereas KH1 and KH2 domains bind eEF1A1 (XREF_SUPPLEMENTARY, right bottom panel)."
"Given that the only difference between KH1to4 IMP1 and KH3 & 4 IMP1 is the presence of KH1 and KH2 domains in the former, it is highly likely that confluentin binds to the KH1 & 2 di-domain to exert its inhibitory effect on IMP1-KRAS RNA interaction."
"As calcium influx is one of the most important mechanisms underlying the effectiveness of CD20 crosslinking function [XREF_BIBR], whether GDC-0980 that abrogated calcium influx antagonized KH-1 in apoptosis was investigated."