A basic peptide within the juxtamembrane region is required for EGF receptor dimerization

Aifa, Sami; Aydin, Jan; Nordvall, Gunnar; Lundström, Ingemar; Svensson, Samuel; Hermanson, Ola

doi:10.1016/j.yexcr.2004.08.032

Cited by 56 publications

(73 citation statements)

References 21 publications

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“…This interaction leads to receptor dimerization and activation as well as to colony growth in soft agar. The ErbB1 juxtamembrane region containing the nuclear localization sequence (NLS), which is important for ErbB1 kinase activation (23), is also important both for nucleolin-ErbB1 interaction and for nucleolin-induced ErbB1 dimerization and activation (24). In addition, we have shown that this effect is mediated by the C-terminal 212 amino acids of nucleolin (24).…”

Section: Introductionmentioning

confidence: 90%

Oncogenic Synergism between ErbB1, Nucleolin, and Mutant Ras

et al. 2011

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Alterations in the ErbB family of growth factor receptors, their signaling components, and mutational activation of Ras proteins are major contributors to malignant transformation. Recently, mutant Ras was shown to be capable of activating ErbB receptors in a ligand-independent manner. Furthermore, it was observed that nucleolin, a transcriptional regulator and ribosome biogenesis factor, can bind both K-Ras and the cytoplasmic tail of ErbB receptors to enhance ErbB receptor activation. However, the functional significance of these interactions to cancer pathogenesis has not been probed. Here, we show that endogenous nucleolin interacts simultaneously in vivo with endogenous Ras and ErbB1 (EGFR) in cancer cells. The C-terminal 212 amino acids of nucleolin were determined to be sufficient to interact with ErbB1 and all Ras protein isoforms (H-, N-, and KRas). Nucleolin partially colocalizes with Ras at the plasma membrane. Moreover, activated but not wild-type Ras facilitates nucleolin interaction with ErbB1 and stabilizes ErbB1 receptor levels. Most importantly, these three oncogenes synergistically facilitate anchorage-independent cell growth in vitro and tumor growth in vivo. Our findings suggest strategies to target nucleolin as a general approach to inhibiting ErbB-and Ras-driven cancers. Cancer Res; 71(6); 2140-51. Ó2011 AACR.

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Section: Introductionmentioning

confidence: 90%

Oncogenic Synergism between ErbB1, Nucleolin, and Mutant Ras

et al. 2011

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“…It has been hypothesized that this sequence may interact with the negatively charged head groups of the lipids in the membrane bilayer, leading to the association of this sequence with the membrane (1,34). By contrast, Aifa et al (35,36) have proposed that a negatively charged stretch of amino acids from residues 979 -991 in the C-terminal tail interacts with the basic residues in the juxtamembrane domain of the EGF receptor. As residues 979 -991 are absent in the cЈ698-and cЈ973-EGF receptors, both of which exhibit negative cooperativity and positive linkage, these negatively charged residues cannot be involved in the allosteric regulation of EGF binding affinity.…”

Section: Discussionmentioning

confidence: 99%

The Intracellular Juxtamembrane Domain of the Epidermal Growth Factor (EGF) Receptor Is Responsible for the Allosteric Regulation of EGF Binding

Macdonald-Obermann

Pike

2009

Journal of Biological Chemistry

115

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We have previously shown that the binding of epidermal growth factor (EGF) to its receptor can best be described by a model that involves negative cooperativity in an aggregating system ( The EGF 2 receptor is a tyrosine kinase composed of an ϳ620-amino-acid extracellular domain that recognizes and binds EGF, a single pass ␣-helical transmembrane domain, and an intracellular tyrosine kinase domain, encompassing roughly residues 685-950 (1). In addition, the receptor contains an ϳ230-amino-acid-long C-terminal tail that contains the bulk of the sites of receptor autophosphorylation (2-4). An intracellular juxtamembrane domain of about 40 residues connects the transmembrane domain to the kinase domain and has been shown to be crucial in the allosteric activation of the EGF receptor kinase (5, 6).In the membrane, the EGF receptor exists as a monomer, but a wealth of data indicate that the binding of EGF induces the formation of EGF receptor dimers (7-10). Dimerization appears to be mediated in large part by the extracellular domain of the receptor, which is comprised of four subdomains, designated I through IV. X-ray crystallography data suggest that in the absence of ligand, the extracellular domain is held in a closed configuration through the interaction of loops or arms that extend from the backs of subdomains II and IV (11). Upon binding of EGF, this intramolecular tether is released, allowing the receptor to adopt an open conformation in which EGF is tightly bound between subdomains I and III. In this configuration, the "dimerization arm" that was previously involved in tethering the receptor closed mediates the formation of a backto-back EGF receptor dimer (12, 13).Analyses of the binding of 125 I-EGF to its receptor have invariably resulted in concave up Scatchard plots that have been interpreted as indicating the presence of two classes of EGF binding sites. However, we have recently used global analysis of the binding of 125 I-EGF to cells expressing increasing levels of EGF receptors to show that EGF binding is best described by a model involving negative cooperativity in an aggregating system (14) (see Fig. 6). Ligand binding is negatively cooperative if the binding of ligand to the first site on a dimer reduces the affinity of the ligand for binding to the second site on the dimer.The concept of cooperativity only applies to existing dimers. It does not relate to the effect of ligand on the assembly or disassembly of those dimers. The effect of ligand on the formation of receptor dimers is captured in the concept of linkage (15,16). If ligand binding is positively linked to dimer formation, then ligand promotes the assembly of receptor dimers. In a monomer-dimer equilibrium, positive linkage arises when a ligand binds with higher affinity to the first site on the dimer than to the monomer. Under these circumstances, the ligand will preferentially bind to the dimer, shifting the equilibrium in favor of the dimeric species. In the case of the EGF receptor, biochemical data suggest that EGF induces recept...

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“…This consensus tripartite NLS seems to be unique for the EGFR family, as it is not present in the above-mentioned receptors. The 13-amino-acid tripartite NLS of the EGFR protein has been shown to be required for receptor dimerization (69). However, other studies showed that mutation of EGFR tripartite NLS or deletion of the ErbB2 tripartite NLS did not affect receptor protein membrane localization and activation of MAPK signaling (13,14).…”

Section: Discussionmentioning

confidence: 99%

Characterization of a Novel Tripartite Nuclear Localization Sequence in the EGFR Family

Hsu

Hung

2007

Journal of Biological Chemistry

217

195

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Aberrant expression of epidermal growth factor receptor (EGFR) is present in many human tumors. Several reports have shown that EGFR is translocated into the nucleus during liver regeneration and in several types of cells and tissues such as placenta and thyroid. Nuclear EGFR is associated with transcription, DNA synthesis, and DNA repair activity and serves as a prognostic marker in breast carcinoma and oropharyngeal squamous cell cancer. However, the nuclear localization sequence (NLS) of EGFR has not been extensively examined. In this study, we have shown that the juxtamembrane region of EGFR harbors a putative NLS with three clusters of basic amino acids (RRRHIVRKRTLRR (amino acids 645-657)) that mediates the nuclear localization of EGFR. We found that this newly characterized tripartite NLS is conserved among the EGFR family members (EGFR, ErbB2, ErbB3, and ErbB4) and is able to move each to the nucleus. Further, this tripartite NLS could also mediate the nuclear localization of other known cytoplasmic proteins such as pyruvate kinase. We have demonstrated that mutating one of the three basic amino acid clusters (R or K3 A) leads to significant impairment of the nuclear localization of EGFR and that of a green fluorescent protein-pyruvate kinase-NLS reporter protein. Our results show that this tripartite NLS is distinct from the traditional mono-and bipartite NLS and reveal a mechanism that could account for the nuclear localization of membrane receptors. The epidermal growth factor receptor (EGFR)2 family of receptor tyrosine kinases (RTKs), which includes EGFR (ErbB1/HER1), ErbB2 (HER2/neu), ErbB3 (HER3), and ErbB4 (HER4), is well known to function as signal transducers at the cell membrane. Classic RTKs contain an extracellular domain, a hydrophobic transmembrane domain, and an intracellular domain. Binding between RTK and their ligands is thought to initiate homodimerization or heterodimerization of the receptor (1); subsequently, dimerized RTKs are activated through their intrinsic tyrosine kinase activities by tyrosine phosphorylation (2, 3). The activated, dimerized RTKs then recruit other signaling molecules to elicit numerous downstream signaling cascades (4 -6) for regulating cellular proliferation, differentiation, and programmed cell death (2, 7).Evidence is emerging to suggest that either full-length or fragmented EGFR family members can be shuttled from the plasma membrane to the nucleus (8 -20). Putative nuclear localization sequences (NLSs) of EGFR family members are thought to be required for the nuclear function of EGFR as a transcriptional co-activator (9, 13). A classical monopartite NLS, RRRRHSP, resembling the NLS for the simian virus 40 (SV40) large T (LT) antigen has been identified in the C terminus of ErbB3 (11). Putative NLSs for ErbB2 and ErbB4 that mediate the nuclear localization of either full-length ErbB2 or truncated ErbB4 have also been identified in the juxtamembrane (JM) region (14, 21). However, whether these putative NLSs within the JM region of EGFR family members coul...

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A basic peptide within the juxtamembrane region is required for EGF receptor dimerization

Cited by 56 publications

References 21 publications

Oncogenic Synergism between ErbB1, Nucleolin, and Mutant Ras

Oncogenic Synergism between ErbB1, Nucleolin, and Mutant Ras

The Intracellular Juxtamembrane Domain of the Epidermal Growth Factor (EGF) Receptor Is Responsible for the Allosteric Regulation of EGF Binding

Characterization of a Novel Tripartite Nuclear Localization Sequence in the EGFR Family

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