Deregulation and mislocalization of the cytokinesis regulator ECT2 activate the Rho signaling pathways leading to malignant transformation.
The human ECT2 protooncogene encodes a guanine nucleotide exchange factor for the Rho GTPases and regulates cytokinesis. Although the oncogenic form of ECT2 contains an N-terminal truncation, it is not clear how the structural abnormality of ECT2 causes malignant transformation. Here we show that both the removal of the negative regulatory domain and alteration of subcellular localization are required to induce the oncogenic activity of ECT2. The transforming activity of oncogenic ECT2 was strongly inhibited by dominant negative Rho GTPases, suggesting the involvement of Rho GTPases in ECT2 transformation. Although deletion of the N-terminal cell cycle regulator-related domain (N) of ECT2 did not activate its transforming activity, removal of the small central domain (S), which contains two nuclear localization signals (NLSs), significantly induced the activity. The ECT2 N domain interacted with the catalytic domain and significantly inhibited the focus formation by oncogenic ECT2. Interestingly, the introduction of the NLS mutations in the S domain of N-terminally truncated ECT2 dramatically induced the transforming activity of this otherwise non-oncogenic derivative. Among the known Rho GTPases expressed in NIH 3T3 cells, RhoA was predominantly activated by oncogenic ECT2 in vivo. Therefore, the mislocalization of structurally altered ECT2 might cause the untimely activation of cytoplasmic Rho GTPases leading to the malignant transformation.
Saito
Shin'ichi
S
Molecular Tumor Biology Section, Basic Research Laboratory, NCI, National Institutes of Health, Bethesda, Maryland 20892-4255, USA.
Liu
Xiu-Fen
XF
Kamijo
Keiju
K
Raziuddin
Razi
R
Tatsumoto
Takashi
T
Okamoto
Isamu
I
Chen
Xiaoyan
X
Lee
Chong-Chou
CC
Lorenzi
Matthew V
MV
Ohara
Naoya
N
Miki
Toru
T
D000199:Actins / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D020134:Catalytic Domain; D002471:Cell Transformation, Neoplastic; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D017353:Gene Deletion; D017930:Genes, Reporter; D005822:Genetic Vectors; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D018715:Microscopy, Video; D008954:Models, Biological; D009154:Mutation; D041681:NIH 3T3 Cells; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis* / Q000502:physiology*; D015398:Signal Transduction; D013997:Time Factors; D015533:Transcriptional Activation; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D000199:Actins; D018076:DNA, Complementary; C489850:ECT2 protein, human; C080509:Ect2 protein, mouse; D011518:Proto-Oncogene Proteins; D008156:Luciferases; D020741:rho GTP-Binding Proteins