A chemical and genetic approach to the mode of action of fumagillin.
Previous mode of action studies identified methionine aminopeptidase 2 (MetAP-2) as the target of the antiangiogenic natural product fumagillin and its drug candidate analog, TNP-470. We report here that TNP-470-mediated MetAP-2 inhibition blocks noncanonical Wnt signaling, which plays a critical role in development, cell differentiation, and tumorigenesis. Consistent with this finding, antisense MetAP-2 morpholino oligonucleotide injection in zebrafish embryos phenocopies gastrulation defects seen in noncanonical Wnt5 loss-of-function zebrafish mutants. MetAP-2 inhibition or depletion blocks signaling downstream of the Wnt receptor Frizzled, but upstream of Calmodulin-dependent Kinase II, RhoA, and c-Jun N-terminal Kinase. Moreover, we demonstrate that TNP-470 does not block the canonical Wnt/beta-catenin pathway. Thus, TNP-470 selectively regulates noncanonical over canonical Wnt signaling and provides a unique means to explore and dissect the biological systems mediated by these pathways.
Zhang
Yi
Y
Department of Molecular, Cell, and Developmental Biology, Yale University, New Haven, Connecticut 06520, USA.
Yeh
Jing Ruey
JR
Mara
Andrew
A
Ju
Rong
R
Hines
John F
JF
Cirone
Pasquale
P
Griesbach
Hilary L
HL
Schneider
Igor
I
Slusarski
Diane C
DC
Holley
Scott A
SA
Crews
Craig M
CM
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D003510:Cyclohexanes / Q000494:pharmacology*; D000072261:Dishevelled Proteins; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism; D005124:Eye Abnormalities / Q000139:chemically induced / Q000235:genetics; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D051157:Frizzled Receptors; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D000077609:O-(Chloroacetylcarbamoyl)fumagillol; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010641:Phenotype; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D043562:Receptors, G-Protein-Coupled; D012717:Sesquiterpenes / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D013623:Tail / Q000002:abnormalities / Q000737:chemistry; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*; D000071818:Wnt-5a Protein; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D020533:Angiogenesis Inhibitors; D003510:Cyclohexanes; D000072261:Dishevelled Proteins; D005231:Fatty Acids, Unsaturated; D051157:Frizzled Receptors; C489743:Fzd2 protein, mouse; D016376:Oligonucleotides, Antisense; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; D012717:Sesquiterpenes; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C480282:Wnt5a protein, zebrafish; D029961:Zebrafish Proteins; C026211:fumagillin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D000626:Aminopeptidases; C571082:methionine aminopeptidase-2, zebrafish; C106541:methionine aminopeptidase 2; D008666:Metalloendopeptidases; D020742:rhoA GTP-Binding Protein; D000077609:O-(Chloroacetylcarbamoyl)fumagillol