Purα Is Essential for Postnatal Brain Development and Developmentally Coupled Cellular Proliferation As Revealed by Genetic Inactivation in the Mouse

Drosophila Pur-α binds to trinucleotide-repeat containing cellular RNAs and translocates to the early oocyte

“…A knockout of Pur-α in mice is viable until shortly after birth, where the mutant animals die from neurological abnormalities such as seizures and tremor. 5 Under certain circumstances, mutant animals can survive for were performed under identical conditions, in many cases the same reaction was analyzed in parallel by EMSA and fluorescence anisotropy and/or microscale temperature gradient thermophoresis. For fluorescence anisotropy, 40 μl of the reactions were transferred into a dark, low-binding surface 96-well plate and measured in a Tecan Infinite M1000 plate-reading fluorescence spectrometer.…”

“…Biochemical analysis of neuronal RNA-transport granules detected the presence of Pur-α as a tightly associated protein that is required for mRNA transport and the progression of transport granules, revealing that Pur-α also functions in the cytoplasm and binds RNA. 4 Deletion of pur-α in mice results in perinatal lethality and abnormal brain morphology, 5,6 consistent with a neuronal requirement of Pur-α. RNase treatment disrupted the transport granules and liberated Pur-α together with its paralog Pur-β, 4 indicating that the mRNP transport complex must be stabilized by RNA.…”

“…Not surprisingly, animal models of these diseases have therefore focused on the analysis of effects in the nervous system, and neuropathological changes are discernible in pur-α knockout mice. 5,11 Analysis of the amino acid sequence revealed that Pur-α contains three occurrences of a characteristic sequence stretch called PUR-repeat; the first two repeats fold together into one nucleic acid binding domain, while the third repeat mediates protein dimerization by forming an intermolecular nucleic acid binding domain with the C-terminal PUR-repeat of a second Pur-α molecule. Crystallographic analysis of Drosophila Pur-α has revealed the three dimensional structure of its nucleic acid binding domain, leading to the identification of amino acids that are essential for nucleic acid binding.…”

A Bifunctional Intronic Element Regulates the Expression of the Arginine/Lysine Transporter Cat-1 via Mechanisms Involving the Purine-rich Element Binding Protein A (Purα)

“…C6 rat glioma cells were cultured in media supplemented with 5% heat-inactivated fetal bovine serum and 5% calf serum. Mouse embryonic fibroblasts (MEFs) with or without homozygous deletions of the Pur␣ gene (15) or the CHOP gene (16) were grown in media supplemented with 10% fetal bovine serum. FuGENE 6 HD (Roche Applied Science) was used to transfect cultured cells according to the manufacturer's instructions.…”

Evidence for the involvement of purα in response to DNA replication stress

“…24 Recent results using Purα knockout mice further support the notion that Purα is essential for cell proliferation during brain development. 25 Purα -/-mice develop neurological problems with severe tremor and spontaneous seizures, presumably due to a lack of neuronal proliferation. 25 However, the mechanism of Purα function in this neuronal defect remains unknown.…”