Calcium ionophore A23187 as a regulator of gene expression in mammalian cells.

Resendez, E; Ting, Jerry; Kim, Kyu Seong; Wooden, Scott K.; Lee, Amy

doi:10.1083/jcb.103.6.2145

Cited by 103 publications

(63 citation statements)

References 35 publications

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“…Several studies implicate HSPs as part of a generalized stress response. Induction of HSP mRNAs after thermal, ischemic, or traumatic injury of the nervous system has been demonstrated for several of the axonally synthesized HSPs that we have identified (Resendez et al, 1986;Lowenstein et al, 1991;Tedeschi and Ciavarra, 1997;Costigan et al, 1998;Liu et al, 1998;Yu et al, 1999;Mengesdorf et al, 2001;Hori et al, 2002). The HSPs are best characterized as molecular chaperones that can facilitate protein folding and formation of heteromolecular complexes and direct proteins to subcellular regions and organelles (Welch, 1992;Ohtsuka and Suzuki, 2000).…”

Section: Axonal Synthesis Of Heat Shock Proteinsmentioning

confidence: 99%

Differential Transport and Local Translation of Cytoskeletal, Injury-Response, and Neurodegeneration Protein mRNAs in Axons

Willis¹,

Li²,

Zheng³

et al. 2005

J. Neurosci.

361

391

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Recent studies have begun to focus on the signals that regulate axonal protein synthesis and the functional significance of localized protein synthesis. However, identification of proteins that are synthesized in mammalian axons has been mainly based on predictions. Here, we used axons purified from cultures of injury-conditioned adult dorsal root ganglion (DRG) neurons and proteomics methodology to identify axonally synthesized proteins. Reverse transcription (RT)-PCR from axonal preparations was used to confirm that the mRNA for each identified protein extended into the DRG axons. Proteins and the encoding mRNAs for the cytoskeletal proteins ␤-actin, peripherin, vimentin, ␥-tropomyosin 3, and cofilin 1 were present in the axonal preparations. In addition to the cytoskeletal elements, several heat shock proteins (HSP27, HSP60, HSP70, grp75, ␣B crystallin), resident endoplasmic reticulum (ER) proteins (calreticulin, grp78/BiP, ERp29), proteins associated with neurodegenerative diseases (ubiquitin C-terminal hydrolase L1, rat ortholog of human DJ-1/Park7, ␥-synuclein, superoxide dismutase 1), anti-oxidant proteins (peroxiredoxins 1 and 6), and metabolic proteins (e.g., phosphoglycerate kinase 1 (PGK 1), ␣ enolase, aldolase C/Zebrin II) were included among the axonally synthesized proteins. Detection of the mRNAs encoding each of the axonally synthesized proteins identified by mass spectrometry in the axonal compartment indicates that the DRG axons have the potential to synthesize a complex population of proteins. Local treatment of the DRG axons with NGF or BDNF increased levels of cytoskeletal mRNAs into the axonal compartment by twofold to fivefold but had no effect on levels of the other axonal mRNAs studied. Neurotrophins selectively increased transport of ␤-actin, peripherin, and vimentin mRNAs from the cell body into the axons rather than changing transcription or mRNA survival in the axonal compartment.

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Section: Axonal Synthesis Of Heat Shock Proteinsmentioning

confidence: 99%

Differential Transport and Local Translation of Cytoskeletal, Injury-Response, and Neurodegeneration Protein mRNAs in Axons

Willis¹,

Li²,

Zheng³

et al. 2005

J. Neurosci.

361

391

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“…Despite the sequence similarity between the mammalian glucose-regulated core sequence and the S. cerevisiae UPRE, no single element in this promoter region appears necessary and sufficient to mediate transcriptional induction as described for the UPRE in yeast cells. In addition, although transcriptional activation in response to conditions that disrupt protein folding in the ER correlates with changes in activities of protein kinases and phosphatases (Resendez et al 1986;Koong et al 1994a;Cao et al 1995;Chen et al 1998), a signaling molecule that responds to unfolded protein in the ER to induce transcription of the ER protein chaperone genes has not been identified. In this report we describe the identification and characterization of a human gene product that is equivalent to Ire1p of S. cerevisiae and functions as a proximal sensor for the UPR in mammalian cells.…”

mentioning

confidence: 99%

A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells

Tirasophon¹,

Welihinda²,

Kaufman³

1998

Genes Dev.

848

727

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Eukaryotes respond to the presence of unfolded protein in the endoplasmic reticulum (ER) by up-regulating the transcription of genes encoding ER protein chaperones, such as BiP. We have isolated a novel human cDNA encoding a homolog to Saccharomyces cerevisiae Ire1p, a proximal sensor for this signal transduction pathway in yeast. The gene product hIre1p is a type 1 transmembrane protein containing a cytoplasmic domain that is highly conserved to the yeast counterpart having a Ser/Thr protein kinase domain and a domain homologous to RNase L. However, the luminal domain has extensively diverged from the yeast gene product. hIre1p expressed in mammalian cells displayed intrinsic autophosphorylation activity and an endoribonuclease activity that cleaved the 5 splice site of yeast HAC1 mRNA, a substrate for the endoribonuclease activity of yeast Ire1p. Overexpressed hIre1p was localized to the ER with particular concentration around the nuclear envelope and some colocalization with the nuclear pore complex. Expression of Ire1p mRNA was autoregulated through a process that required a functional hIre1p kinase activity. Finally, overexpression of wild-type hIre1p constitutively activated a reporter gene under transcriptional control of the rat BiP promoter, whereas expression of a catalytically inactive hIre1p acted in a trans-dominant-negative manner to prevent transcriptional activation of the BiP promoter in response to ER stress induced by inhibition of N-linked glycosylation. These results demonstrate that hIre1p is an essential proximal sensor of the unfolded protein response pathway in mammalian cells.

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“…First, the transcriptional activation of the GRP78 gene is sensitive to the protein synthesis inhibitor cycloheximide (8,20), whereas the induction of the heat-inducible HSP70 gene does not require de novo protein synthesis. In fact, it has been demonstrated that the heat shock regulatory factor can bind to the heat shock element in the presence of cycloheximide (26).…”

mentioning

confidence: 99%

“…To facilitate the search for cellular factors which interact with the DNA regulatory domain, we utilized the HeLa cell system, which can be grown in suspension to provide high yields of cellular extracts. We have shown previously that GRP78 transcripts can be detected in a variety of mammalian cell lines (20). Since the focus of this study was on the utilization of the human HeLa S3 cell line as a source of functional protein extracts, we first investigated the expression of the endogenous GRP78 gene in these cells.…”

mentioning

confidence: 99%

“…For control and calcium ionophore (A23187) treatment, cells were placed in fresh medium 8 h before cytoplasmic RNA extraction; for calcium ionophore treatment, cells were incubated for 5 h with the ionophore before RNA extraction; and for the medium starvation condition, the cells were kept in the original medium and incubated for an additional 24 h before RNA extraction. The cytoplasmic RNA samples (5 ,ug) were electrophoresed on a formaldehyde denaturing gel, trans-blotted onto a nitrocellulose filter, and hybridized with radiolabeled hamster GRP78 cDNA plasmid p3C5 (23a) as described previously (10,20). The arrow indicates the position of the 2.7-kilobase GRP78 transcript.…”

mentioning

confidence: 99%

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Identification of highly conserved regulatory domains and protein-binding sites in the promoters of the rat and human genes encoding the stress-inducible 78-kilodalton glucose-regulated protein.

1988

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The gene encoding GRP78 has been shown to be constitutively expressed in many cell types and is inducible by the calcium ionophore A23187. To understand the regulation of GRP78 transcription, we analyzed the components that control its basal-level expression. By transfecting deletions into cells, we have identified a 54-nucleotide cis-acting regulatory element important for high basal-level expression and a contiguous 50-nucleotide element for both basal-level expression and A23187 induction. Using DNase footprinting assays with both rat and human GRP78 promoters, we demonstrated that the protein factors present in the HeLa cell nuclear extracts bind to the regulatory regions identified by the deletion studies. This domain contains a palindromic sequence and is highly conserved among GRP genes in Caenorhabditis elegans, chicks, rats, and humans.

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Calcium ionophore A23187 as a regulator of gene expression in mammalian cells.

Cited by 103 publications

References 35 publications

Differential Transport and Local Translation of Cytoskeletal, Injury-Response, and Neurodegeneration Protein mRNAs in Axons

Differential Transport and Local Translation of Cytoskeletal, Injury-Response, and Neurodegeneration Protein mRNAs in Axons

A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells

Identification of highly conserved regulatory domains and protein-binding sites in the promoters of the rat and human genes encoding the stress-inducible 78-kilodalton glucose-regulated protein.

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