Alternative splicing of the N-terminal cytosolic and transmembrane domains of P2X7 controls gating of the ion channel by ADP-ribosylation.
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Alternative splicing of the N-terminal cytosolic and transmembrane domains of P2X7 controls gating of the ion channel by ADP-ribosylation. / Schwarz, Nicole; Drouot, Laurent; Nicke, Annette; Fliegert, Ralf; Boyer, Olivier; Guse, Andreas H.; Haag, Friedrich; Adriouch, Sahil; Koch-Nolte, Friedrich.
In: PLOS ONE, Vol. 7, No. 7, 7, 2012, p. 41269.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Alternative splicing of the N-terminal cytosolic and transmembrane domains of P2X7 controls gating of the ion channel by ADP-ribosylation.
AU - Schwarz, Nicole
AU - Drouot, Laurent
AU - Nicke, Annette
AU - Fliegert, Ralf
AU - Boyer, Olivier
AU - Guse, Andreas H.
AU - Haag, Friedrich
AU - Adriouch, Sahil
AU - Koch-Nolte, Friedrich
PY - 2012
Y1 - 2012
N2 - P2X7 is a homotrimeric ion channel with two transmembrane domains and a large extracellular ATP-binding domain. It plays a key role in the response of immune cells to danger signals released from cells at sites of inflammation. Gating of murine P2X7 can be induced by the soluble ligand ATP, as well as by NAD(+)-dependent ADP-ribosylation of arginine 125, a posttranslational protein modification catalyzed by the toxin-related ecto-enzymes ART2.1 and ART2.2. R125 is located at the edge of the ligand-binding crevice. Recently, an alternative splice variant of P2X7, designated P2X7(k), was discovered that differs from the previously described variant P2X7(a) in the N-terminal 42 amino acid residues composing the first cytosolic domain and most of the Tm1 domain. Here we compare the two splice variants of murine P2X7 with respect to their sensitivities to gating by ADP-ribosylation in transfected HEK cells. Our results show that the P2X7(k) variant is sensitive to activation by ADP-ribosylation whereas the P2X7(a) variant is insensitive, despite higher cell surface expression levels. Interestingly, a single point mutation (R276K) renders the P2X7(a) variant sensitive to activation by ADP-ribosylation. Residue 276 is located at the interface of neighboring subunits approximately halfway between the ADP-ribosylation site and the transmembrane domains. Moreover, we show that naive and regulatory T cells preferentially express the more sensitive P2X7(k) variant, while macrophages preferentially express the P2X7(a) variant. Our results indicate that differential splicing of alternative exons encoding the N-terminal cytosolic and transmembrane domains of P2X7 control the sensitivity of different immune cells to extracellular NAD(+) and ATP.
AB - P2X7 is a homotrimeric ion channel with two transmembrane domains and a large extracellular ATP-binding domain. It plays a key role in the response of immune cells to danger signals released from cells at sites of inflammation. Gating of murine P2X7 can be induced by the soluble ligand ATP, as well as by NAD(+)-dependent ADP-ribosylation of arginine 125, a posttranslational protein modification catalyzed by the toxin-related ecto-enzymes ART2.1 and ART2.2. R125 is located at the edge of the ligand-binding crevice. Recently, an alternative splice variant of P2X7, designated P2X7(k), was discovered that differs from the previously described variant P2X7(a) in the N-terminal 42 amino acid residues composing the first cytosolic domain and most of the Tm1 domain. Here we compare the two splice variants of murine P2X7 with respect to their sensitivities to gating by ADP-ribosylation in transfected HEK cells. Our results show that the P2X7(k) variant is sensitive to activation by ADP-ribosylation whereas the P2X7(a) variant is insensitive, despite higher cell surface expression levels. Interestingly, a single point mutation (R276K) renders the P2X7(a) variant sensitive to activation by ADP-ribosylation. Residue 276 is located at the interface of neighboring subunits approximately halfway between the ADP-ribosylation site and the transmembrane domains. Moreover, we show that naive and regulatory T cells preferentially express the more sensitive P2X7(k) variant, while macrophages preferentially express the P2X7(a) variant. Our results indicate that differential splicing of alternative exons encoding the N-terminal cytosolic and transmembrane domains of P2X7 control the sensitivity of different immune cells to extracellular NAD(+) and ATP.
KW - Animals
KW - Humans
KW - Mice
KW - Mice, Inbred BALB C
KW - Protein Structure, Tertiary
KW - HEK293 Cells
KW - Protein Isoforms/genetics/metabolism
KW - Adenosine Diphosphate Ribose/genetics/metabolism
KW - Adenosine Triphosphate/genetics/metabolism
KW - Alternative Splicing/physiology
KW - Ion Channel Gating/physiology
KW - NAD/genetics/metabolism
KW - Protein Processing, Post-Translational/physiology
KW - Receptors, Purinergic P2X7/genetics/metabolism
KW - T-Lymphocytes, Regulatory/cytology/metabolism
KW - Animals
KW - Humans
KW - Mice
KW - Mice, Inbred BALB C
KW - Protein Structure, Tertiary
KW - HEK293 Cells
KW - Protein Isoforms/genetics/metabolism
KW - Adenosine Diphosphate Ribose/genetics/metabolism
KW - Adenosine Triphosphate/genetics/metabolism
KW - Alternative Splicing/physiology
KW - Ion Channel Gating/physiology
KW - NAD/genetics/metabolism
KW - Protein Processing, Post-Translational/physiology
KW - Receptors, Purinergic P2X7/genetics/metabolism
KW - T-Lymphocytes, Regulatory/cytology/metabolism
U2 - 10.1371/journal.pone.0041269
DO - 10.1371/journal.pone.0041269
M3 - SCORING: Journal article
VL - 7
SP - 41269
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 7
M1 - 7
ER -
