Pax6-induced alteration of cell fate: Shape changes, expression of neuronal α tubulin, postmitotic phenotype, and cell migration

Cartier, Laetitia; Laforge, Térèse; Féki, Anis; Arnaudeau, Serge; Dubois‐Dauphin, Michel; Krause, Karl‐Heinz

doi:10.1002/neu.20225

Cited by 28 publications

(27 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[19][20][21] Accordingly, we previously observed that ectopic Pax6 expression provoked a postmitotic G 0 /G 1 cell-cycle arrest in HeLa cells. 9 This was however not altered by hRFPL1 RNA interference (data not shown), suggesting that hRFPL1 influence on cell-cycle progression is not sufficient to produce cell-cycle exit and differentiation in Pax6-expressing cells.…”

Section: Discussionmentioning

confidence: 88%

“…Given the possible role of hRFPL1 on cell-cycle and in silico prediction of putative p53 binding sites on its promoter, we examined the influence of p53 on hRFPL1 transcriptional regulation. Upon Pax6-elicited hRFPL1 expression and p53 activation in HeLa cells, 4,9 we observed using chromatin immunoprecipitation (ChIP) that p53 interacted in vivo with hRFPL1 promoter (Figure 1a). After blocking p53 activity using either cyclic-Pifithrin-a, an inhibitor of p53 nuclear translocation 10 or a p53 shRNA, 11,12 we observed that Pax6-induced hRFPL1 expression was significantly reduced (Figure 1b).…”

Section: Resultsmentioning

confidence: 96%

See 1 more Smart Citation

Primate-specific RFPL1 gene controls cell-cycle progression through cyclin B1/Cdc2 degradation

et al. 2010

View full text Add to dashboard Cite

Ret finger protein-like 1 (RFPL1) is a primate-specific target gene of Pax6, a key transcription factor for pancreas, eye and neocortex development. However, its cellular activity remains elusive. In this article, we report that Pax6-elicited expression of the human (h)RFPL1 gene in HeLa cells can be enhanced by in vivo p53 binding to its promoter and therefore investigated the hypothesis that hRFPL1 regulates cell-cycle progression. Upon expression in these cells, hRFPL1 decreased cell number through a kinase-dependent mechanism as PKC activates and Cdc2 inhibits hRFPL1 activity. hRFPL1 antiproliferative activity led to an increased cell population in G 2 /M phase and specific cyclin B1 and Cdc2 downregulations, which were precluded by a proteasome inhibitor. Specifically, cytoplasm-localized hRFPL1 prevented cyclin B1 and Cdc2 accumulation during interphase. Consequently, cells showed a delayed entry into mitosis and cell-cycle lengthening resulting from a threefold increase in G 2 phase duration. Given previous reports that RFPL1 is expressed during cell differentiation, its impact on cell-cycle lengthening therefore provides novel insights into primate-specific development. The human Ret finger protein-like (hRFPL)1,2,3 genes (OMIM 605968, 605969, 605970) are recently identified targets of Pax6, which is notably a key transcription factor for pancreas, eye and neocortex development. [1][2][3] In this line, high hRFPL1,2,3 expression is found at the onset of neurogenesis in differentiating embryonic stem cells and in the developing neocortex. 4 In addition, the study of RFPL1,2,3 evolutionary history revealed that they are only found in Old World monkeys and great apes and show features believed to be important for human brain evolution. 4 Yet, the cellular activity of RFPL1,2,3 is still unknown. A murine RFPL (mRFPL) protein, encoded by an ancestral gene not belonging to the RFPL1,2,3 gene subfamily, 4 has also been cloned. 5 Previously reported as being expressed only in testis, ovaries and oocytes, 5,6 mRFPL has been shown to interact with the destruction box motif of cyclin B1 -the Cdc2 activating partner for driving germ cells through metaphases I and II 7 -and with proteins of the proteasome, speculating on mRFPL ability to elicit cyclin B1 degradation to control meiosis progression. 6 However, the fact that cyclin B1 and Cdc2 also form a key complex for controlling cell entry into mitosis 8 and our recent observations that the RFPL genes are also expressed in tissues in which cells divide mitotically 4 suggest that the RFPL proteins could regulate other aspects of cell division.We therefore examined RFPL-mediated control of mitotic cell-cycle progression by focusing on hRFPL1. Because no endogenously hRFPL1-expressing cell type suitable for this kind of study has been reported to date, we examined the influence of hRFPL1 gain of function on HeLa cells, a reference cell system for examining cell-cycle regulation. We report that hRFPL1 is an antiproliferative gene that controls G 2 -M phase transition, ...

show abstract

Section: Discussionmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 96%

Primate-specific RFPL1 gene controls cell-cycle progression through cyclin B1/Cdc2 degradation

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Electroporation of R-cadherin into Pax6-deficient embryos rescued correct outgrowth of pioneer axons during early brain patterning in the mouse (Andrews and Mastick, 2003). Overexpression of Pax6 in transfected HeLa cells induces changes in cellular shape and migration, prompts massive expression of neuronal atubulin, and establishes a postmitotic phenotype (Cartier et al, 2006). Moreover, it was shown that Pax6 regulates the transcription of cadherin 4 (Liu et al, 2001), L1-CAM (Chalepakis et al, 1994;Meech et al, 1999), NCAM (Holst et al, 1997) and a4integrin (Zaniolo et al, 2004).…”

Section: Introductionmentioning

confidence: 97%

Retinal patterning by Pax6‐dependent cell adhesion molecules

Rungger‐Brändle¹,

Ripperger

Steiner

et al. 2010

Developmental Neurobiology

View full text Add to dashboard Cite

Long-standing evidence gained from Pax6 mutant embryos pointed to an involvement of Pax6-dependent cell adhesion molecules in patterning the central nervous system and, in particular, the retina. However, direct evidence for such pathways remained elusive. We here present direct evidence that knockdown of Pax6 expression by morpholino antisense molecules in Xenopus embryos and knockdown of maternal N-cadherin (mNcad), N-cadherin (Ncad) and neural cell adhesion molecule (NCAM) produce similar phenotypes. Eye formation is reduced and retinal lamination is heavily disorganized. In Pax6 knockdown embryos, the levels of mRNAs coding for these cell adhesion molecules are markedly reduced. Overexpression of Pax6 efficiently rescues the phenotype of Pax6 knockdown embryos and restores expression of these putative target genes. Rescue of Pax6-deficiency by the putative target gene mNcad moderately rescues eye formation. The promoters of the genes coding for cell adhesion molecules contain several putative Pax6 binding sites, as determined by computer analysis. Chromatin immunoprecipitation shows that, in embryonic heads, Pax6 binds to promoter regions containing such predicted binding sites. Thus, several cell adhesion molecules are direct target genes of Pax6 and cooperate in retinal patterning.

show abstract

“…Myocytes can be differentiated to osteogenic lineage by transfection of RunX1/Cbfa1 [84]. Recently, HeLa tumor cells were transdifferentiated to neurons by forcing the expression of the Pax-6 transcription factor [85]. In vivo, liver cells can be made to differentiate to pancreatic f cells by codelivery of Neuro-D and β-cellulin using gutless adenovirus and thus treat diabetes in an animal model [86].…”

Section: Transdifferentiationmentioning

confidence: 98%

“…For safety purposes, long-term studies on larger animals are necessary in order to predict whether or not a given therapeutic approach will be efficient and without side effects in humans. [16,[20][21][22] Express BMP-7 and HGF [23,73] Immunogenicity Downregulation of MHC class I and co-receptor molecules siRNA, anti-sense RNA, express CTLA-4 Ig [34,110] Autoimmune disease TNF, pro-inflammatory cytokines Express soluble proinflammatory cytokine receptors [110] Express anti-inflammatory cytokines [110] Control unwanted growth Pro-drug activating enzymes Expressing HSV-tk [5] Direct differentiation to a particular pathway Transcription factors Such as Brachyury, Myo-D, Nurr-1, RunX1/Cbfa-1, Pax-6, Neuro-D and β-cellulin [77][78][79][80][81][82][83][84][85][86] Growth/differentiation factors Such as BMP-2,-4, TGF-β, IGF-1 and GDNF [87][88][89][90][91] BMP: Bone morphogenetic protein; IGF: Insulin growth factor; siRNA: Small interfering RNA; TGF:Transforming growth factor.…”

Section: Future Perspectivementioning

confidence: 99%

Engineering Stem Cells for Therapy

et al. 2006

View full text Add to dashboard Cite

The differentiation of a stem cell is dependent on the environmental cues that it receives and can be modulated by the expression of different master regulators or by secreted factors or inducers. The use of genetically modified stem cells to express the required factors can direct differentiation along the requisite pathway. This approach to the engineering of stem cells is important, as control of the pluripotentiality of stem cells is necessary in order to avoid unwanted growth, migration or differentiation to nontarget tissues. The authors provide an overview of the stem cell engineering field, highlighting challenges and solutions, and focusing on recent developments in therapeutic applications in areas such as autoimmunity, CNS lesions, bone and joint diseases, cancer and myocardial infarction.

show abstract

Pax6-induced alteration of cell fate: Shape changes, expression of neuronal α tubulin, postmitotic phenotype, and cell migration

Cited by 28 publications

References 68 publications

Primate-specific RFPL1 gene controls cell-cycle progression through cyclin B1/Cdc2 degradation

Primate-specific RFPL1 gene controls cell-cycle progression through cyclin B1/Cdc2 degradation

Retinal patterning by Pax6‐dependent cell adhesion molecules

Engineering Stem Cells for Therapy

Contact Info

Product

Resources

About