Erratum FGF-mediated mesoderm induction involves the Src-family kinase Laloo

Weinstein, Daniel C.; Marden, Jennifer J.; Carnevali, Francesca; Hemmati‐Brivanlou, Ali

doi:10.1038/27706

Cited by 382 publications

(93 citation statements)

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“…The earliest onset of zygotic expression was detected for src mRNA, confirming the results of Collett and Steele (1992). The laloo temporal expression profile presented here complements the partial laloo expression data previously published by Weinstein et al (1998) and also correlates very well with the RT-PCR expression analysis described in Song et al (2001). In order to quantitatively compare the expression efficiency of individual SFK and csk genes, we normalised their expression levels to that of lyn mRNA (Fig.…”

Section: Quantitative Real-time Pcrsupporting

confidence: 87%

Expression patterns of Src-family tyrosine kinases during Xenopus laevis development

Ferjentsik

Šindelka²,

Jonák³

2009

Int. J. Dev. Biol.

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Src family tyrosine kinases (SFKs) play important roles in cell morphology, differentiation, motility and proliferation. Elevated expression and/or specific activity of Src kinases are characteristic for several types of human cancer. However, little information is available about the role and spatio-temporal expression of SFKs in early embryonic development. In this study we characterized, in Xenopus laevis, the expression patterns of five SFK genes src, fyn, yes, lyn and laloo as well as of the csk gene, a negative regulator of SFKs, using RT-qPCR and in situ hybridisation. We found that transcripts of all SFKs and csk were already detectable in one-cell embryos and their levels similarly oscillated during subsequent development. First, after stage 8, the levels of SFK and csk mRNAs began to decrease, reached a minimum between stages 10 and 28 and increased again. In the later stages (33-45), the levels of fyn, yes and csk mRNAs returned to approximately maternal ones, whereas the src, laloo and lyn mRNA transcripts exceeded, up to about 3.5-6-fold, their maternal levels. In situ hybridisation analysis located the SFK and csk transcripts in the animal hemisphere of Xenopus embryos. Subsequent gastrula stages showed signals in ectodermal cells, mid-neurula stage embryos at neural folds, and the tailbud stages showed strong signals in the brain and neural tube. RT-qPCR concentration profiling along the animal-vegetal axis proved in blastula and gastrula the preferential localisation of yes, src, lyn and csk transcripts towards the animal pole in a gradient-like manner. In contrast, laloo and fyn displayed a vegetal pole preference. KEY WORDS: Xenopus laevis, quantitative real-time PCR, in situ hybridisation, early developmentSrc and Src-family protein-tyrosine kinases (SFKs) are protooncogenes and represent one of the nine presently recognised classes of non-receptor tyrosine-kinases (Pellicena and Miller, 2002). As documented in a great many reports the members of the SFK family participate in a variety of signalling pathways that control cell behavior, including differentiation and transformation (for a review see e.g. Blume-Jensen and Hunter, 2001). On the other hand, the role of SFK members in developmental processes has been examined much less extensively. The experiments carried out on mice demonstrated that Src/Fyn and Src/Yesdouble knockouts die perinatally (Stein et al., 1994) and Src/Fyn/ Yes-triple knockouts at an early stage of embryonic development (Klinghoffer et al., 1999).In frogs Xenopus laevis, Steele and co-workers reported that src, yes, and fyn transcripts were already present in the maternal RNA pool (Steele, 1985;Steele et al., 1989Steele et al., , 1990. In contrast to Int.

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Section: Quantitative Real-time Pcrsupporting

confidence: 87%

Expression patterns of Src-family tyrosine kinases during Xenopus laevis development

Ferjentsik

Šindelka²,

Jonák³

2009

Int. J. Dev. Biol.

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“…Second, growth factor-dependent Spry2 tyrosine phosphorylation was blocked by two Src kinase inhibitors, as well as by a dominant-negative Src mutant. Src and related kinases mediate growth factor signaling (Twamley-Stein et al, 1993;Roche et al, 1995;Kuo et al, 1997;Weinstein et al, 1998) and play important roles in many of the processes affected by Spry overexpression, such as cell proliferation (Impagnatiello et al, 2001), migration (Yigzaw et al, 2001), and neurite outgrowth (Gross et al, 2001). We were unable to demonstrate phosphorylation of Spry1/2 by recombinant Src in vitro and did not consistently detect Spry2 in a complex with Src.…”

Section: Discussionmentioning

confidence: 67%

Tyrosine Phosphorylation of Sprouty Proteins Regulates Their Ability to Inhibit Growth Factor Signaling: A Dual Feedback Loop

Mason

Morrison

Bassit

et al. 2004

MBoC

116

144

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Sprouty proteins are recently identified receptor tyrosine kinase (RTK) inhibitors potentially involved in many developmental processes. Here, we report that Sprouty proteins become tyrosine phosphorylated after growth factor treatment. We identified Tyr55 as a key residue for Sprouty2 phosphorylation and showed that phosphorylation was required for Sprouty2 to inhibit RTK signaling, because a mutant Sprouty2 lacking Tyr55 augmented signaling. We found that tyrosine phosphorylation of Sprouty2 affected neither its subcellular localization nor its interaction with Grb2, FRS2/SNT, or other Sprouty proteins. In contrast, Sprouty2 tyrosine phosphorylation was necessary for its binding to the Src homology 2-like domain of c-Cbl after fibroblast growth factor (FGF) stimulation. To determine whether c-Cbl was required for Sprouty2-dependent cellular events, Sprouty2 was introduced into c-Cbl-wild-type and -null fibroblasts. Sprouty2 efficiently inhibited FGF-induced phosphorylation of extracellular signal-regulated kinase 1/2 in c-Cbl-null fibroblasts, thus indicating that the FGF-dependent binding of c-Cbl to Sprouty2 was dispensable for its inhibitory activity. However, c-Cbl mediates polyubiquitylation/proteasomal degradation of Sprouty2 in response to FGF. Last, using Src-family pharmacological inhibitors and dominant-negative Src, we showed that a Src-like kinase was required for tyrosine phosphorylation of Sprouty2 by growth factors. Thus, these data highlight a novel negative and positive regulatory loop that allows for the controlled, homeostatic inhibition of RTK signaling.

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“…Microinjection, explant dissection, and culture were performed as described (Hemmati-Brivanlou and Melton, 1994). RT-PCR was performed as described (Wilson and Hemmati-Brivanlou, 1995;Weinstein et al, 1998). Primers constructed for this study are as follows: ␣-Globin (top), 5'-TTGCTCTGTGG-GGCAAAATC; ␣-Globin (bottom), 5'-GGTTGTAGGCATGCAGGTCA; Endodermin (top), 5'-CTCGCTCTGG-ACAAAACTC; Endodermin (bottom), 5'-GAGGTTGCTGATGGGAATG; GA-TA-5 (top), 5'-GCCAGACGAATACAC-CTACAG; GATA-5 (bottom), 5'-AGAA-ACCCAAGATACCACCAT; XlHbox8 (top), 5'-CCTACAGCAACCCCTTG-GTA; XlHbox8 (bottom), 5'-GGGCTCTT-GTGTAGGCTGTC; IFABP (top), 5'-GC-CTTTGATGGAACTTGGAA; IFABP (bottom), 5'-CTGTAGGAACCAGGCAC-CAT.…”

Section: Xenopus Embryo Assaysmentioning

confidence: 99%

Mouse Mix gene is activated early during differentiation of ES and F9 stem cells and induces endoderm in frog embryos

Mohn

Chen

Dias

et al. 2003

Developmental Dynamics

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In frog and zebrafish, the Mix/Bix family of paired type homeodomain proteins play key roles in specification and differentiation of mesendoderm. However, in mouse, only a single Mix gene (mMix) has been identified to date and its function is unknown. We have analyzed the expression of mouse Mix RNA and protein in embryos, embryoid bodies formed from embryonic stem cells and F9 teratocarcinoma cells, as well as several differentiated cell types. Expression in embryoid bodies in culture mirrors that in embryos, where Mix is transcribed transiently in primitive (visceral) endoderm (VE) and in nascent mesoderm. In F9 cells induced by retinoic acid to differentiate to VE, mMix is coordinately expressed with three other endodermal transcription factors, well before AFP, and its protein product is localized to the nucleus. In a subpopulation of nascent mesodermal cells from embryonic stem cell embryoid bodies, mMix is coexpressed with Brachyury. Intriguingly, mMix mRNA is detected in a population (T؉Flk1؉) of cells which may contain hemangioblasts, before the onset of hematopoiesis and activation of hematopoietic markers. In vitro and in vivo, mMix expression in nascent mesoderm is rapidly down-regulated and becomes undetectable in differentiated cell types. In the region of the developing gut, mMix expression is confined to the mesoderm of midand hindgut but is absent from definitive endoderm. Injection of mouse mMix RNA into early frog embryos results in axial truncation of developing tadpoles and, in animal cap assays, mMix alone is sufficient to activate expression of several endodermal (but not mesodermal) markers. Although these observations do not exclude a possible cellautonomous function for mMix in mesendodermal progenitor cells, they do suggest an additional, non-cell autonomous role in nascent mesoderm in the formation and/or patterning of adjacent definitive endoderm. Developmental Dynamics 226:446 -459, 2003.

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Erratum FGF-mediated mesoderm induction involves the Src-family kinase Laloo

Cited by 382 publications

References 1 publication

Expression patterns of Src-family tyrosine kinases during Xenopus laevis development

Expression patterns of Src-family tyrosine kinases during Xenopus laevis development

Tyrosine Phosphorylation of Sprouty Proteins Regulates Their Ability to Inhibit Growth Factor Signaling: A Dual Feedback Loop

Mouse Mix gene is activated early during differentiation of ES and F9 stem cells and induces endoderm in frog embryos

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