Impaired Binding of 14-3-3 to C-RAF in Noonan Syndrome Suggests New Approaches in Diseases with Increased Ras Signaling

Molzan, Manuela; Schumacher, Benjamin; Ottmann, C.; Baljuls, Angela; Polzien, Lisa; Weyand, Michael; Thiel, Philipp; Rose, Rolf; Rose, Micheline; Kuhenne, Philipp; Kaiser, Markus; Rapp, Ulf R.; Kuhlmann, Jürgen; Ottmann, C.

doi:10.1128/mcb.01636-09

Cited by 113 publications

(101 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, the salt bridge between pNth1 1-751 residues R81 and D85 is responsible for an abrupt change in the direction of the polypeptide chain (Figs. 2B and 3B), thus again mimicking the role of the Pro residue at the +2 position (9,25,27,28). The intervening linker sequence between the two 14-3-3 binding motifs of pNth1 (residues 64−79) is ordered compared with known structures of 14-3-3 protein complexes, most likely a result of numerous interactions with helices A1 and A3 of Bmh1 (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…3). Interactions between pNth1 1-751 residues D63 and N64 at the +3 and +4 positions and Bmh1 residues K51 and N52 appear to force a change in the direction of the polypeptide chain, mimicking the role of the Pro residue, which is frequently found in 14-3-3 binding motifs at the +2 position relative to the phosphorylated residue (9,(25)(26)(27)(28). The second 14-3-3 binding motif of pNth1 (sequence RRGpS 83 ED) also does not contain a Pro residue at the +2 position relative to the phosphorylated residue (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…2B). As a result of these interactions, the C-terminal portions of both pNth1 motifs leave the binding grooves in similar orientation as polypeptide chains of other proteins and peptides including, for example, AANAT (9), the heat shock protein beta-6 (27), and short phosphopeptides (25,28), that were cocrystalized with 14-3-3 and whose 14-3-3 binding motifs contain proline residue at the +2 position (Fig. 3).…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Alblova

Smidova

Dočekal

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The 14-3-3 proteins, a family of highly conserved scaffolding proteins ubiquitously expressed in all eukaryotic cells, interact with and regulate the function of several hundreds of partner proteins. Yeast neutral trehalases (Nth), enzymes responsible for the hydrolysis of trehalose to glucose, compared with trehalases from other organisms, possess distinct structure and regulation involving phosphorylation at multiple sites followed by binding to the 14-3-3 protein. Here we report the crystal structures of yeast Nth1 and its complex with Bmh1 (yeast 14-3-3 isoform), which, together with mutational and fluorescence studies, indicate that the binding of Nth1 by 14-3-3 triggers Nth1’s activity by enabling the proper 3D configuration of Nth1’s catalytic and calcium-binding domains relative to each other, thus stabilizing the flexible part of the active site required for catalysis. The presented structure of the Bmh1:Nth1 complex highlights the ability of 14-3-3 to modulate the structure of a multidomain binding partner and to function as an allosteric effector. Furthermore, comparison of the Bmh1:Nth1 complex structure with those of 14-3-3:serotonin N-acetyltransferase and 14-3-3:heat shock protein beta-6 complexes revealed similarities in the 3D structures of bound partner proteins, suggesting the highly conserved nature of 14-3-3 affects the structures of many client proteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Alblova

Smidova

Dočekal

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Therefore, DUF1669 was required for CRAF binding, AIG, and the increased phosphorylation of ERK1/2 compared with control cells (Figure 5, B, C, and H, and Figure 7H). Regulation of CRAF kinase activity involves the binding of 14-3-3 proteins, which maintain CRAF in an inactive conformation in the cytoplasm (11,12). Displacement of 14-3-3 proteins allows CRAF translocation to the plasma membrane and subsequent activation of its kinase activity, resulting in the phosphorylation and activation of downstream MAPK proteins MEK and ERK (11,12).…”

Section: Figurementioning

confidence: 99%

FAM83B mediates EGFR- and RAS-driven oncogenic transformation

et al. 2012

View full text Add to dashboard Cite

Aberrant regulation of growth signaling is a hallmark of cancer development that often occurs through the constitutive activation of growth factor receptors or their downstream effectors. Using validation-based insertional mutagenesis (VBIM), we identified family with sequence similarity 83, member B (FAM83B), based on its ability to substitute for RAS in the transformation of immortalized human mammary epithelial cells (HMECs). We found that FAM83B coprecipitated with a downstream effector of RAS, CRAF. Binding of FAM83B with CRAF disrupted CRAF/14-3-3 interactions and increased CRAF membrane localization, resulting in elevated MAPK and mammalian target of rapamycin (mTOR) signaling. Ablation of FAM83B inhibited the proliferation and malignant phenotype of tumor-derived cells or RAS-transformed HMECs, implicating FAM83B as a key intermediary in EGFR/RAS/MAPK signaling. Analysis of human tumor specimens revealed that FAM83B expression was significantly elevated in cancer and was associated with specific cancer subtypes, increased tumor grade, and decreased overall survival. Cumulatively, these results suggest that FAM83B is an oncogene and potentially represents a new target for therapeutic intervention. IntroductionThe discovery of targets suitable for the development of specific and effective anticancer therapies remains one of the principal challenges facing cancer research. The identification of genes involved in tumorigenesis is essential for devising new targeted therapeutics and can be greatly facilitated by phenotypic-based forward genetic screens for mutations contributing to malignant transformation in human cell models. We recently created a validation-based insertional mutagenesis (VBIM) strategy that expands the application of reversible promoter insertion to nearly any type of mammalian cell (1). The VBIM strategy uses the unique transcriptomes of different human epithelial cell types and provides opportunities for the identification of tissue-specific oncogenes and tumor suppressors. The VBIM lentiviruses alter the unique transcriptome of the model system by introducing promoters into the genome, resulting in dominant genetic alterations that increase the expression of sequences neighboring the insertion sites. By using Cre recombinase-mediated excision of the VBIM promoter, one can revert the VBIM-specific mutants and distinguish them from spontaneous mutants, allowing spontaneous mutants to be eliminated from further study.We have used the VBIM strategy to identify family with sequence similarity 83, member B (FAM83B), as a putative oncogene capable of promoting the transformation of immortalized human mammary epithelial cells (HMECs). We demonstrated that elevated FAM83B expression stimulated aberrant activation of MAPK signaling by altering binding of regulatory 14-3-3 proteins to CRAF and increasing CRAF membrane localization. In addition to driving cellular transformation, FAM83B mRNA was significantly elevated in many human tumor tissues. Ablation of FAM83B from breast cancer cells with ele...

show abstract

“…Thereby, they modify the target proteins' ability to interact with other proteins, enzymatic activity or subcellular localization [21]. For example, 14-3-3 proteins regulate the activity of the cellcycle phosphatase Cdc25 [22,23], the protein kinase C-RAF [24,25] and the transcriptional modulator YAP [26,27], and stabilize the tumour suppressor p53 [28,29]. 14-3-3 proteins have been associated with various cancers [30], the virulence of human pathogenic organisms [31,32], and the development of neurodegenerative diseases [33].…”

Section: Introductionmentioning

confidence: 99%

Subcellular localization of full-length human myeloid leukemia factor 1 (MLF1) is independent of 14-3-3 proteins

Molzan

Ottmann²

2013

Cellular and Molecular Biology Letters

Self Cite

View full text Add to dashboard Cite

Myeloid leukemia factor 1 (MLF1) is associated with the development of leukemic diseases such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). However, information on the physiological function of MLF1 is limited and mostly derived from studies identifying MLF1 interaction partners like CSN3, MLF1IP, MADM, Manp and the 14-3-3 proteins. The 14-3-3-binding site surrounding S34 is one of the only known functional features of the MLF1 sequence, along with one nuclear export sequence (NES) and two nuclear localization sequences (NLS). It was recently shown that the subcellular localization of mouse MLF1 is dependent on 14-3-3 proteins. Based on these findings, we investigated whether the subcellular localization of human MLF1 was also directly 14-3-3-dependent. Live cell imaging with GFP-fused human MLF1 was used to study the effects of mutations and deletions on its subcellular localization. Surprisingly, we found that the subcellular localization of full-length human MLF1 is 14-3-3-independent, and is probably regulated by other as-yet-unknown proteins.

show abstract

Impaired Binding of 14-3-3 to C-RAF in Noonan Syndrome Suggests New Approaches in Diseases with Increased Ras Signaling

Cited by 113 publications

References 58 publications

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

FAM83B mediates EGFR- and RAS-driven oncogenic transformation

Subcellular localization of full-length human myeloid leukemia factor 1 (MLF1) is independent of 14-3-3 proteins

Contact Info

Product

Resources

About