Inhibition of hematopoietic development from embryonic stem cells by antisense vav RNA.

Wulf, Gerbug M.; Adra, Chaker N.; Lim, Bing

doi:10.1002/j.1460-2075.1993.tb06200.x

Cited by 56 publications

(35 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A random hexamer-primed cDNA strand was synthesized with mouse mammary leukemia virus RT. The RT products were used as templates for PCR amplification by using gene-specific primers (35,64,68). The amount of cDNA synthesized was calibrated by using the relative expression level of hypoxanthine phosphoribosyltransferase (HPRT) as a standard.…”

Section: Methodsmentioning

confidence: 99%

“…The results described above point to the participation of Shp-2 in controlling the development of the myeloerythroid lineages. To corroborate the in vitro colony assays, we examined the expression of several genes that serve as markers of hematopoietic cells by RT-PCR analysis (35,64,68). Total RNAs of wild-type and mutant origins were isolated from ES cells and from EBs collected at selected days.…”

Section: Downloaded Frommentioning

confidence: 99%

See 1 more Smart Citation

A Deletion Mutation in the SH2-N Domain of Shp-2 Severely Suppresses Hematopoietic Cell Development

Shi

Shen

et al. 1997

Molecular and Cellular Biology

158

148

View full text Add to dashboard Cite

Shp-1 and Shp-2 are cytoplasmic protein tyrosine phosphatases that contain two Src homology 2 (SH2) domains. A negative regulatory role of Shp-1 in hematopoiesis has been strongly implicated by the phenotype of motheaten mice with a mutation in the Shp-1 locus, which is characterized by leukocyte hypersensitivity, deregulated mast cell function, and excessive erythropoiesis. A targeted deletion of 65 amino acids in the N-terminal SH2 (SH2-N) domain of Shp-2 leads to an embryonic lethality at midgestation in homozygous mutant mice. To further dissect the Shp-2 function in hematopoietic development, we have isolated homozygous Shp-2 mutant embryonic stem (ES) cells. Significantly reduced hematopoietic activity was observed when the mutant ES cells were allowed to differentiate into embryoid bodies (EBs), compared to the wild-type and heterozygous ES cells. Further analysis of ES cell differentiation in vitro showed that mutation in the Shp-2 locus severely suppressed the development of primitive and definitive erythroid progenitors and completely blocked the production of progenitor cells for granulocytes-macrophages and mast cells. Reverse transcriptase PCR analysis of the mutant EBs revealed reduced expression of several specific marker genes that are induced during blood cell differentiation. Stem cell factor induction of mitogen-activated protein kinase activity was also blocked in Shp-2 mutant cells. Taken together, these results indicate that Shp-2 is an essential component and primarily plays a positive role in signaling pathways that mediate hematopoiesis in mammals. Furthermore, stimulation of its catalytic activity is not sufficient, while interaction via the SH2 domains with the targets or regulators is necessary for its biological functions in cells. The in vitro ES cell differentiation assay can be used as a biological tool in dissecting cytoplasmic signaling pathways.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Downloaded Frommentioning

confidence: 99%

A Deletion Mutation in the SH2-N Domain of Shp-2 Severely Suppresses Hematopoietic Cell Development

Shi

Shen

et al. 1997

Molecular and Cellular Biology

158

148

View full text Add to dashboard Cite

show abstract

“…In this regard, one molecule of potential interest is the 95-kDa product of the Vav protooncogene, another SH2 domain-containing protein which, like PTP1C, has been identified in all hemopoietic lineages and implicated by several lines of evidence in the control of hemopoietic cell growth and differentiation (18 -20). Inhibition of Vav expression, for example, interferes with development of hemopoietic cells from embryonic stem cells (21), and, as is consistent with its participation in a broad range of hemopoietic cell signaling pathways, Vav has been shown to become tyrosine-phosphorylated following cross-linking of antigen receptors on lymphocytes (22)(23)(24), Fc␥ and ⑀ receptors on monocytes and mast cells, respectively (24,25), and c-kit receptors on multiple hemopoietic lineages (26,27). Vav contains a number of structural motifs found in many signaling effectors, including an SH2, a pleckstrin homology, and two SH3 do-* This work was supported in part by grants from the Medical Research Council of Canada and the National Cancer Institute of Canada (NCIC) and by the Health Canada Bureau of Drug Research.…”

mentioning

confidence: 86%

The Tyrosine Phosphatase PTP1C Associates with Vav, Grb2, and mSos1 in Hematopoietic Cells

Kon-Kozlowski

Pani

Pawson

et al. 1996

Journal of Biological Chemistry

114

View full text Add to dashboard Cite

The association of the murine motheaten phenotype of severe hemopoietic dysregulation with loss of PTP1C tyrosine phosphatase activity indicates a critical role for this SH2 domain-containing phosphotyrosine phosphatase in the regulation of hemopoietic cell growth and differentiation. To explore the molecular basis for PTP1C effects on hematopoiesis, we have investigated the possibility that this enzyme interacts with the product of the Vav proto-oncogene, a putative guanine nucleotide exchange factor expressed exclusively in hemopoietic cells. Our data indicate that PTP1C physically associates with Vav in murine spleen cells and in EL4 T lymphoma and P815 mastocytoma cells, and that this interaction is increased following mitogenic stimulation and the induction of both PTP1C and Vav tyrosine phosphorylation. The results also reveal tyrosine phosphatase activity to be present in Vav immunoprecipitates from stimulated splenic and P815 cells and suggest that a major portion of total cellular PTP1C catalytic activity is associated with Vav. As Vav-associated tyrosine phosphatase activity was not detected in PTP1C-deficient motheaten splenic cells, it appears that PTP1C accounts for most, if not all, Vav-coprecipitable tyrosine phosphatase activity in normal cells. The data also demonstrate the capacity of the Vav SH2 domain alone to bind to PTP1C in activated P815 cells, but suggest a role for the two Vav SH3 domains in enhancing this interaction. In addition, the results reveal PTP1C association with two other molecules implicated in Ras activation, the Grb2 adaptor protein and mSos1, a GTP/GDP exchanger for Ras. PTP1C therefore has the capacity to bind and potentially modulate various signaling effectors involved in activation of Ras or Ras-related proteins, and, accordingly, regulation of Ras activation represents a possible mechanism whereby PTP1C influences hemopoietic cellular responses.Among the phosphotyrosine phosphatases (PTP) 1 identified to date, the cytosolic enzyme PTP1C is distinguished by its predominant expression in hemopoietic cells and the presence of two N-terminal located Src homology 2 (SH2) domains, a motif found in only two other PTPs, Syp (PTP1D/SHPTP2) and the Drosophila csw protein (1-6). These properties, together with the recent data linking PTP1C gene mutations to the profound hemopoietic dysregulation manifested by motheaten (me) and viable motheaten (me v ) mice (7-9), reveal a critical role for PTP1C in modulating hemopoietic cell differentiation and growth. As this PTP has been shown to associate with the activated c-kit, erythropoietin, and IL-3 receptors (10 -12) and, more recently, with the B cell antigen receptor complex and the CD22 and Fc␥RIIB1 receptors on lymphocytes (13-15), PTP1C appears to subserve its regulatory role, at least in part, by modulating the signaling capacities of membrane growth factor/antigen/cytokine receptors. As is consistent with the marked overexpansion of multiple hemopoietic cell types observed in PTP1C-deficient motheaten mice, the data concernin...

show abstract

“…The phosphorylation of Vav has been observed after stimulation of the T-cell receptor (Gulbins et al 1993), membrane immunoglobulin (Gulbins et al 1994c), Fc receptor (Margolis et al 1992), interferon receptor (Platanias & Sweet 1994), c-kit (Alai et al 1992), PRLr (Clevenger et al 1995b), and the receptors for IL-1 (Gulbins et al 1994a) and -2 (Evans et al 1993). Antisense experiments have revealed that the inhibition of Vav expression in embryonic stem cells prevents their in vitro differentiation into mature hematopoietic elements (Wulf et al 1993). These findings, however, have been contradicted in a recent study using vav / mouse embryonic stem cells (Zhang et al 1994), as normal hematopoietic differentiation was observed in this system in vitro.…”

Section: Mediators Of Prl-induced Lymphocyte Proliferation -Vavmentioning

confidence: 99%

Prolactin receptor signal transduction in cells of the immune system

Freier²,

Kline³

1998

Journal of Endocrinology

117

View full text Add to dashboard Cite

Inhibition of hematopoietic development from embryonic stem cells by antisense vav RNA.

Cited by 56 publications

References 33 publications

A Deletion Mutation in the SH2-N Domain of Shp-2 Severely Suppresses Hematopoietic Cell Development

A Deletion Mutation in the SH2-N Domain of Shp-2 Severely Suppresses Hematopoietic Cell Development

The Tyrosine Phosphatase PTP1C Associates with Vav, Grb2, and mSos1 in Hematopoietic Cells

Prolactin receptor signal transduction in cells of the immune system

Contact Info

Product

Resources

About