Identification of an invasion-inducing gene, Tiam-1, that encodes a protein with homology to GDP-GTP exchangers for Rho-like proteins

Habets, Gaston; Scholtes, Ellen H.M.; Zuydgeest, David; Kammen, Rob A. van der; Stam, Jord C.; Berns, Anton; Collard, John G.

doi:10.1016/0092-8674(94)90216-x

Cited by 495 publications

(420 citation statements)

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“…Rho GTPases such as Rac1 become activated when bound GDP is exchanged for GTP by a process catalyzed by GEFs such as Tiam1 (Habets et al 1994). A Tiam1 transcript has been detected previously in breast cancer cells (Habets et al 1995).…”

Section: Resultsmentioning

confidence: 99%

“…For example, recent studies have shown that certain metastasis-specific molecules (e.g., CD44v 3,8–10 isoform [Bourguignon et al 1998b, Bourguignon et al 1999] and its associated matrix metalloproteinase, MMP-9 [Bourguignon et al, 1998b; Yu and Stamenkovic 1999], as well as Rho kinase [Bourguignon et al 1999]) are closely associated with the cytoskeleton during tumor cell function. To further examine the regulatory mechanism(s) involved in cytoskeleton-mediated oncogenic signaling leading to tumor cell invasion and migration, we have focused on GEFs (the Dbl or DH family), such as Tiam1, which are known to display oncogenic capability and function as upstream activators of Rho-like GTPases (e.g., Rac1 or Cdc42; Woods et al, 1991; Habets et al 1994; Michiels et al 1995; Nobes and Hall 1995; Van Leeuwen et al 1995). In breast tumor cells, such as SP-1 cells, Tiam1 is detected as a 200-kD protein (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…One of these GEFs is Tiam1 (T-lymphoma invasion and metastasis 1), which was identified by retroviral insertional mutagenesis and selected for its invasive cell behavior in vitro (Habets et al 1994, Habets et al 1995). This molecule is largely hydrophilic and contains several functional domains found in signal transduction proteins.…”

Section: Introductionmentioning

confidence: 99%

“…This molecule is largely hydrophilic and contains several functional domains found in signal transduction proteins. For example, the COOH-terminal region of the Tiam1 molecule has a Dbl homology (DH) domain (Hart et al 1991, Hart et al 1994; Habets et al 1994) and an adjacent pleckstrin homology (PH) domain, which exists in most GEFs (Hart et al 1991, Hart et al 1994; Habets et al 1994; Lemmon et al 1996). In particular, the DH domain of these proteins exhibits GDP/GTP exchange activity for specific members of the Ras superfamily of GTP-binding proteins (Hart et al 1991, Hart et al 1994).…”

Section: Introductionmentioning

confidence: 99%

“…In particular, the DH domain of these proteins exhibits GDP/GTP exchange activity for specific members of the Ras superfamily of GTP-binding proteins (Hart et al 1991, Hart et al 1994). Tiam1 also contains an additional PH domain, a Discs-large homology region (DHR; Habets et al 1994; Pontings and Phillips 1995), and a potential myristoylation site in the NH 2 -terminal part of the protein (Habets et al 1994). …”

Section: Introductionmentioning

confidence: 99%

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Ankyrin–Tiam1 Interaction Promotes Rac1 Signaling and Metastatic Breast Tumor Cell Invasion and Migration

Bourguignon

Zhu

Shao

et al. 2000

The Journal of Cell Biology

116

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Tiam1 (T-lymphoma invasion and metastasis 1) is one of the known guanine nucleotide (GDP/GTP) exchange factors (GEFs) for Rho GTPases (e.g., Rac1) and is expressed in breast tumor cells (e.g., SP-1 cell line). Immunoprecipitation and immunoblot analyses indicate that Tiam1 and the cytoskeletal protein, ankyrin, are physically associated as a complex in vivo. In particular, the ankyrin repeat domain (ARD) of ankyrin is responsible for Tiam1 binding. Biochemical studies and deletion mutation analyses indicate that the 11–amino acid sequence between amino acids 717 and 727 of Tiam1 (717GEGTDAVKRS727L) is the ankyrin-binding domain. Most importantly, ankyrin binding to Tiam1 activates GDP/GTP exchange on Rho GTPases (e.g., Rac1).Using an Escherichia coli–derived calmodulin-binding peptide (CBP)–tagged recombinant Tiam1 (amino acids 393–728) fragment that contains the ankyrin-binding domain, we have detected a specific binding interaction between the Tiam1 (amino acids 393–738) fragment and ankyrin in vitro. This Tiam1 fragment also acts as a potent competitive inhibitor for Tiam1 binding to ankyrin. Transfection of SP-1 cell with Tiam1 cDNAs stimulates all of the following: (1) Tiam1–ankyrin association in the membrane projection; (2) Rac1 activation; and (3) breast tumor cell invasion and migration. Cotransfection of SP1 cells with green fluorescent protein (GFP)–tagged Tiam1 fragment cDNA and Tiam1 cDNA effectively blocks Tiam1–ankyrin colocalization in the cell membrane, and inhibits GDP/GTP exchange on Rac1 by ankyrin-associated Tiam1 and tumor-specific phenotypes. These findings suggest that ankyrin–Tiam1 interaction plays a pivotal role in regulating Rac1 signaling and cytoskeleton function required for oncogenic signaling and metastatic breast tumor cell progression.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Ankyrin–Tiam1 Interaction Promotes Rac1 Signaling and Metastatic Breast Tumor Cell Invasion and Migration

Bourguignon

Zhu

Shao

et al. 2000

The Journal of Cell Biology

116

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Efficiency assessment of the gene trap approach

Voss¹,

Thomas²,

Gruß³

1998

Dev. Dyn.

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The trapping of genes in murine embryonic stem (ES) cells offers three features in one experimental approach: 1) analysis of the expression patterns of unknown genes by using a simple staining method, 2) rapid cloning of unknown genes, and 3) generation of mutant mouse lines. We performed a gene trap screen aimed at the discovery of new genes regulating embryonic development. We have processed 209 gene trap events for expression patterns in chimeric murine embryos. Randomly tested, β‐galactosidase‐positive ES cell clones resulted in vivo in 35% gene trap events showing no β‐galactosidase activity, 39% gene trap events with ubiquitous β‐galactosidase activity, and 26% gene trap events showing β‐galactosidase activity restricted to specific cell types or organs. In vitro preselection reduced gene trap events with ubiquitous β‐galactosidase activity to 10% and increased the gene trap events with restricted β‐galactosidase activity to 64%, making the screening procedure for genes expressed in a restricted manner 2.5‐fold more efficient. In five of the seven gene trap insertions into genes in which the expression pattern during embryogenesis was known, the β‐galactosidase marker gene reproduced faithfully the expression pattern of the trapped gene. 5′‐Rapid amplification of cDNA ends (5′‐RACE) of 28 gene trap events revealed 19 novel mouse genes, 8 known mouse genes, and 1 random trans‐splicing event. Twelve of the 25 mouse lines that crossed to homozygosity showed overt abnormalities. The genomic structure was investigated in four of these gene trap events, which caused obvious abnormalities. In all four cases, the splice‐acceptor gene trap construct was inserted into an exon. One of the 13 gene trap events that did not result in overt abnormalities was examined for the presence of wild‐type mRNA. Homozygous animals were found to produce normal levels of wild‐type mRNA. Evidently, gene trapping does not always provide all three of the features mentioned above. In this paper, we discuss the efficiency of gene trapping and ways in which some problems may be overcome. Dev. Dyn. 1998;212:171–180. © 1998 Wiley‐Liss, Inc.

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A BALB/c 3T3-transformed cell line suitable for transfection assay of metastasis-inducing genes

et al. 1997

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A clonal cell line, 1-1ras1000, transformed by the activated c-Ha-ras oncogene, does not form metastases after i.v. injection into mice (experimental metastasis assay). Here, we show that this cell line is useful as a recipient to detect metastasis-inducing genes, using a transfection assay. Cells (1-1ras1000) were susceptible to metastasis induction by transfection with either v-src or genomic DNA from a v-src-and v-fos-transferred highly metastatic rat cell line (SR202). The susceptibility of 1-1ras1000 cells for lung metastasis induction was suitable for a genomic transfection assay to detect a metastasis-inducing gene in the transfected cells which had incorporated genomic DNA from donor meta-static tumor cells. When DNAs extracted from 7 human tumors were tested for metastasis induction, 2 DNAs from nonmalignant tumors (non-tumorigenic tumors in athymic nude mice) (2/2) were negative and 4 DNAs from malignant tumors (4/5) were positive in 1-1ras1000 cells for primary transfection. In one of the resulting metastases, the ability to metastasize was also transferred in the second and third cycles of genomic DNA transfection at high frequencies. All of the resulting metastases carried the human repetitive Alu sequence. Neither rearrangements of the endogenous c-Ha-ras nor changes of protein amounts were detected. Recipient 1-1ras1000 cells had a negligible rate of spontaneously meta-static conversion during in vitro cultivation and transfection processes. The resulting metastasized cells were easily isolated from the lung after culturing in selection medium containing G418 (geneticin). Isolated cells stably retained the ability to form metastatic lung nodules when re-injected into mice. Thus, 1-1ras1000 cells appear to be a useful system for the isolation of metastasis-inducing genes from human meta-static tumors. Int. J. Cancer, 71:88-93, 1997. r 1997 Wiley-Liss, Inc. Most primary tumors arising in the body are thought to have no ability to spread invasively. In these tumors, at the final stages of development and progression, a metastatic subpopulation of tumor cells that can form discontinuous tumors at sites near to or distant from the primary site may appear. Such a cell population, with metastatic potential, is distinct from the primary tumorigenic cell population (Fidler and Hart, 1982; Talmadge and Fidler, 1982; Schirrmacher, 1985). One paradigm about tumor progression proposes that a finite number of non-random chromosomal alterations and genetic changes accumulate and are responsible for each step of multiple-stage carcinogenesis (Vogelstein et al., 1988), including metastasis (Dear and Kefford, 1990; Liotta et al., 1991). The metastatic conversion of non-metastatic tumor cells appears to be controlled at 2 levels of genetic activity. The first is where genes act dominantly to induce metastasis, e.g., variant type CD44 (Gunthert et al., 1991) and TIAM1 (Habets et al., 1994). The second is where genes act as metastasis suppressors. Loss of the genes predisposes metastatic conversion from the non-metast...

show abstract

Identification of an invasion-inducing gene, Tiam-1, that encodes a protein with homology to GDP-GTP exchangers for Rho-like proteins

Cited by 495 publications

References 34 publications

Ankyrin–Tiam1 Interaction Promotes Rac1 Signaling and Metastatic Breast Tumor Cell Invasion and Migration

Ankyrin–Tiam1 Interaction Promotes Rac1 Signaling and Metastatic Breast Tumor Cell Invasion and Migration

Efficiency assessment of the gene trap approach

A BALB/c 3T3-transformed cell line suitable for transfection assay of metastasis-inducing genes

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