CDP-diacylglycerol synthetase-controlled phosphoinositide availability limits VEGFA signaling and vascular morphogenesis

Pan, Weijun; Pham, Van N.; Stratman, Amber N.; Castranova, Daniel; Kamei, Makoto; Kidd, Kameha R.; Lo, Brigid D.; Shaw, Kenna Mills; Torres-Vázquez, Jesús; Mikelis, Constantinos M.; Gutkind, J. Silvio; Davis, George E.; Weinstein, Brant M.

doi:10.1182/blood-2012-02-408328

Cited by 39 publications

(46 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…6H). After we conducted our confirmation experiments, another group published a paper showing that a different mutation in the same gene gave a similar phenotype (Pan et al 2012). Together, these experiments indicate that MMAPPR correctly identified cds2 as the causative mutant in the zy14 line.…”

Section: Validationmentioning

confidence: 72%

MMAPPR: Mutation Mapping Analysis Pipeline for Pooled RNA-seq

et al. 2013

View full text Add to dashboard Cite

Forward genetic screens in model organisms are vital for identifying novel genes essential for developmental or disease processes. One drawback of these screens is the labor-intensive and sometimes inconclusive process of mapping the causative mutation. To leverage high-throughput techniques to improve this mapping process, we have developed a Mutation Mapping Analysis Pipeline for Pooled RNA-seq (MMAPPR) that works without parental strain information or requiring a preexisting SNP map of the organism, and adapts to differential recombination frequencies across the genome. MMAPPR accommodates the considerable amount of noise in RNA-seq data sets, calculates allelic frequency by Euclidean distance followed by Loess regression analysis, identifies the region where the mutation lies, and generates a list of putative coding region mutations in the linked genomic segment. MMAPPR can exploit RNA-seq data sets from isolated tissues or whole organisms that are used for gene expression and transcriptome analysis in novel mutants. We tested MMAPPR on two known mutant lines in zebrafish, nkx2.5 and tbx1, and used it to map two novel ENU-induced cardiovascular mutants, with mutations found in the ctr9 and cds2 genes. MMAPPR can be directly applied to other model organisms, such as Drosophila and Caenorhabditis elegans, that are amenable to both forward genetic screens and pooled RNA-seq experiments. Thus, MMAPPR is a rapid, cost-efficient, and highly automated pipeline, available to perform mutant mapping in any organism with a well-assembled genome.

show abstract

Section: Validationmentioning

confidence: 72%

MMAPPR: Mutation Mapping Analysis Pipeline for Pooled RNA-seq

et al. 2013

View full text Add to dashboard Cite

show abstract

“…A photoreceptor-specific isoform of Drosophila CDS was shown to be necessary for the maintenance of PtdIns(4,5)P 2 levels and hence for a sustained light response, and CDS mutant flies developed light-dependent retinal degeneration (1736). Recent studies also identified mutations in one of the CDS genes in zebrafish causing specific defects in blood vessel formation and angiogenesis, and these effects were attributed to the rundown of PtdIns(4,5)P 2 levels in the VEGF-stimulated endothelial cells (1188). These studies also concluded that CDS enzyme activities are necessary for maintaining the signaling pool of PtdIns(4,5)P 2 .…”

Section: Phosphatidylinositol Synthesismentioning

confidence: 99%

Phosphoinositides: Tiny Lipids With Giant Impact on Cell Regulation

Balla

2013

Physiological Reviews

1,396

1,655

View full text Add to dashboard Cite

Phosphoinositides (PIs) make up only a small fraction of cellular phospholipids, yet they control almost all aspects of a cell's life and death. These lipids gained tremendous research interest as plasma membrane signaling molecules when discovered in the 1970s and 1980s. Research in the last 15 years has added a wide range of biological processes regulated by PIs, turning these lipids into one of the most universal signaling entities in eukaryotic cells. PIs control organelle biology by regulating vesicular trafficking, but they also modulate lipid distribution and metabolism via their close relationship with lipid transfer proteins. PIs regulate ion channels, pumps, and transporters and control both endocytic and exocytic processes. The nuclear phosphoinositides have grown from being an epiphenomenon to a research area of its own. As expected from such pleiotropic regulators, derangements of phosphoinositide metabolism are responsible for a number of human diseases ranging from rare genetic disorders to the most common ones such as cancer, obesity, and diabetes. Moreover, it is increasingly evident that a number of infectious agents hijack the PI regulatory systems of host cells for their intracellular movements, replication, and assembly. As a result, PI converting enzymes began to be noticed by pharmaceutical companies as potential therapeutic targets. This review is an attempt to give an overview of this enormous research field focusing on major developments in diverse areas of basic science linked to cellular physiology and disease.

show abstract

“…The dose of injected morpholinos/mRNA: 16 ng for Fbw7 splicing, Fbw7 605 and Fbw7 544 MO; 8 ng for KLF2a MO; and 0.1 ng for hFBW7. Confocal images were obtained using an Olympus Fluoview 1000 confocal microscope and data were analyzed as described [59]. All the experiments were repeated at least three times, and only representative results were shown.…”

Section: Fish Care Morpholinos Microinjection and Whole-mount Mrna mentioning

confidence: 99%

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

Wang

Liu

et al. 2013

Cell Res

Self Cite

View full text Add to dashboard Cite

F-box and WD repeat domain-containing 7 (FBW7), the substrate-binding subunit of E3 ubiquitin ligase SCF FBW7 (a complex of SKP1, cullin-1 and FBW7), plays important roles in various physiological and pathological processes. Although FBW7 is required for vascular development, its function in the endothelium remains to be investigated. In this study, we show that FBW7 is an important regulator of endothelial functions, including angiogenesis, leukocyte adhesion and the endothelial barrier integrity. Using RNA interference, we found that the depletion of FBW7 markedly impairs angiogenesis in vitro and in vivo. We identified the zinc finger transcription factor Krüppel-like factor 2 (KLF2) as a physiological target of FBW7 in endothelial cells. Knockdown of FBW7 expression resulted in the accumulation of endogenous KLF2 protein in endothelial cells. FBW7-mediated KLF2 destruction was shown to depend on the phosphorylation of KLF2 via glycogen synthase kinase-3 (GSK3) at two conserved phosphodegrons. Mutating these phosphodegron motifs abolished the FBW7-mediated degradation and ubiquitination of KLF2. The siRNAmediated knockdown of FBW7 showed that KLF2 is an essential target of FBW7 in the regulation of endothelial functions. Moreover, FBW7-mediated KLF2 degradation was shown to be critical for angiogenesis in teratomas and in zebrafish development. Taken together, our study suggests a role for FBW7 in the processes of endothelial cell migration, angiogenesis, inflammation and barrier integrity, and provides novel insights into the regulation of KLF2 stability in vivo.

show abstract

CDP-diacylglycerol synthetase-controlled phosphoinositide availability limits VEGFA signaling and vascular morphogenesis

Cited by 39 publications

References 58 publications

MMAPPR: Mutation Mapping Analysis Pipeline for Pooled RNA-seq

MMAPPR: Mutation Mapping Analysis Pipeline for Pooled RNA-seq

Phosphoinositides: Tiny Lipids With Giant Impact on Cell Regulation

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

Contact Info

Product

Resources

About