Phospholipase C-γ is required for agonist-induced Ca2+ entry

Randen L. Patterson; Damian B. Van Rossum; Diana L. Ford; Kenneth J. Hurt; Sun Sik Bae; Pann Ghill Suh; Tomohiro Kurosaki; Solomon H. Snyder; Donald L. Gill

doi:10.1016/S0092-8674(02)01045-0

Phospholipase C-γ is required for agonist-induced Ca²⁺ entry

Randen L. Patterson, Damian B. Van Rossum, Diana L. Ford, Kenneth J. Hurt, Sun Sik Bae, Pann Ghill Suh, Tomohiro Kurosaki, Solomon H. Snyder, Donald L. Gill

School of Medicine

Research output: Contribution to journal › Article › peer-review

164 Scopus citations

Abstract

We report here that PLC-γ isoforms are required for agonist-induced Ca²⁺ entry (ACE). Overexpressed wild-type PLC-γ1 or a lipase-inactive mutant PLC-γ1 each augmented ACE in PC12 cells, while a deletion mutant lacking the region containing the SH3 domain of PLC-γ1 was ineffective. RNA interference to deplete either PLC-γ1 or PLC-γ2 in PC12 and A7r5 cells inhibited ACE. In DT40 B lymphocytes expressing only PLC-γ2, overexpressed muscarinic M5 receptors (M5R) activated ACE. Using DT40 PLC-γ2 knockout cells, M5R stimulation of ER Ca²⁺ store release was unaffected, but ACE was abolished. Normal ACE was restored by transient expression of PLC-γ2 or a lipase-inactive PLC-γ2 mutant. The results indicate a lipase-independent role of PLC-γ in the physiological agonist-induced activation of Ca²⁺ entry.

Original language	English (US)
Pages (from-to)	529-541
Number of pages	13
Journal	Cell
Volume	111
Issue number	4
DOIs	https://doi.org/10.1016/S0092-8674(02)01045-0
State	Published - Nov 15 2002

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/S0092-8674(02)01045-0

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title = "Phospholipase C-γ is required for agonist-induced Ca2+ entry",

abstract = "We report here that PLC-γ isoforms are required for agonist-induced Ca2+ entry (ACE). Overexpressed wild-type PLC-γ1 or a lipase-inactive mutant PLC-γ1 each augmented ACE in PC12 cells, while a deletion mutant lacking the region containing the SH3 domain of PLC-γ1 was ineffective. RNA interference to deplete either PLC-γ1 or PLC-γ2 in PC12 and A7r5 cells inhibited ACE. In DT40 B lymphocytes expressing only PLC-γ2, overexpressed muscarinic M5 receptors (M5R) activated ACE. Using DT40 PLC-γ2 knockout cells, M5R stimulation of ER Ca2+ store release was unaffected, but ACE was abolished. Normal ACE was restored by transient expression of PLC-γ2 or a lipase-inactive PLC-γ2 mutant. The results indicate a lipase-independent role of PLC-γ in the physiological agonist-induced activation of Ca2+ entry.",

author = "Patterson, {Randen L.} and {Van Rossum}, {Damian B.} and Ford, {Diana L.} and Hurt, {Kenneth J.} and Bae, {Sun Sik} and Suh, {Pann Ghill} and Tomohiro Kurosaki and Snyder, {Solomon H.} and Gill, {Donald L.}",

note = "Funding Information: We thank Dr. Kirill Kiselyov, Dr. Darren F. Boehning, and Dr. Robert E. Rothe for useful discussion, Kartik Venkatachalam for technical assistance, Dr. Craig Montell for the kind gift of TRPC3 antibody, and Dr. Lutz Birnbaumer for the kind gift of the HEK T3-65 cell line. Special thanks go to Barbara van Rossum, Mary Allen, Thomas Magnetti, and Morgen Patterson for their support of all things WAC. Research was supported by USPHS grant MH-18501 and Research Scientist Award DA-00074 (S.H.S.) and by NIH grants NH65090 (R.L.P.) and HL55426 (D.L.G.)",

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doi = "10.1016/S0092-8674(02)01045-0",

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T1 - Phospholipase C-γ is required for agonist-induced Ca2+ entry

AU - Patterson, Randen L.

AU - Van Rossum, Damian B.

AU - Ford, Diana L.

AU - Hurt, Kenneth J.

AU - Bae, Sun Sik

AU - Suh, Pann Ghill

AU - Kurosaki, Tomohiro

AU - Snyder, Solomon H.

AU - Gill, Donald L.

N1 - Funding Information: We thank Dr. Kirill Kiselyov, Dr. Darren F. Boehning, and Dr. Robert E. Rothe for useful discussion, Kartik Venkatachalam for technical assistance, Dr. Craig Montell for the kind gift of TRPC3 antibody, and Dr. Lutz Birnbaumer for the kind gift of the HEK T3-65 cell line. Special thanks go to Barbara van Rossum, Mary Allen, Thomas Magnetti, and Morgen Patterson for their support of all things WAC. Research was supported by USPHS grant MH-18501 and Research Scientist Award DA-00074 (S.H.S.) and by NIH grants NH65090 (R.L.P.) and HL55426 (D.L.G.)

PY - 2002/11/15

Y1 - 2002/11/15

N2 - We report here that PLC-γ isoforms are required for agonist-induced Ca2+ entry (ACE). Overexpressed wild-type PLC-γ1 or a lipase-inactive mutant PLC-γ1 each augmented ACE in PC12 cells, while a deletion mutant lacking the region containing the SH3 domain of PLC-γ1 was ineffective. RNA interference to deplete either PLC-γ1 or PLC-γ2 in PC12 and A7r5 cells inhibited ACE. In DT40 B lymphocytes expressing only PLC-γ2, overexpressed muscarinic M5 receptors (M5R) activated ACE. Using DT40 PLC-γ2 knockout cells, M5R stimulation of ER Ca2+ store release was unaffected, but ACE was abolished. Normal ACE was restored by transient expression of PLC-γ2 or a lipase-inactive PLC-γ2 mutant. The results indicate a lipase-independent role of PLC-γ in the physiological agonist-induced activation of Ca2+ entry.

AB - We report here that PLC-γ isoforms are required for agonist-induced Ca2+ entry (ACE). Overexpressed wild-type PLC-γ1 or a lipase-inactive mutant PLC-γ1 each augmented ACE in PC12 cells, while a deletion mutant lacking the region containing the SH3 domain of PLC-γ1 was ineffective. RNA interference to deplete either PLC-γ1 or PLC-γ2 in PC12 and A7r5 cells inhibited ACE. In DT40 B lymphocytes expressing only PLC-γ2, overexpressed muscarinic M5 receptors (M5R) activated ACE. Using DT40 PLC-γ2 knockout cells, M5R stimulation of ER Ca2+ store release was unaffected, but ACE was abolished. Normal ACE was restored by transient expression of PLC-γ2 or a lipase-inactive PLC-γ2 mutant. The results indicate a lipase-independent role of PLC-γ in the physiological agonist-induced activation of Ca2+ entry.

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JO - Cell

JF - Cell

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Phospholipase C-γ is required for agonist-induced Ca²⁺ entry

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