Overexpression of SH2-SH2-SH3 domain of phospholipase C-γ1 blocks PDGF-induced inositol phosphate generation in NIH 3T3 cells

Jong Soo Chang, Do Sik Min, Sun Sik Bae, Jae Ho Kim, Young Han Lee, Sung Ho Ryu, Pann Ghill Suh

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Src homology (SH) 2 and 3 domains are known to be binding motifs for protein-protein interaction in signaling molecules. Among several PLC isozymes, only PLC-γ contains the SH domain between the X and Y domains, which are known to have catalytic activity. To elucidate the functional roles of the SH2-SH2-SH3 domain of PLC-γ1 in cellular signaling, we constructed a truncated cDNA encoding the SH2-SH2-SH3 domain of PLC-γ1 (p60SH2/SH3) and expressed it in NIH 3T3 cells. Cells expressing p60SH2/SH3 did not show any change in cell shape no oncogenesity. Even though in a serum depleted condition, a portion of p60SH2/SH3 existed as constitutively phosphorylated on its tyrosine residues. Furthermore, cells expressing p60SH2/SH3 did not respond to PDGF-induced IPs formation whereas vector transfected control cells showed dose-dependent IPs generation upon PDGF stimulation. The tyrosine phosphorylation level of endogenous PLC-γ1 by PDGF, however, was comparable to that of the control cells. On the other hand, IPs accumulation by PLC-β activation occurred to a comparable level. Taken together, p60SH2/SH3 molecules selectively inhibited the IPs accumulation catalyzed by PLC-γ1. This result suggests that the SH2-SH2-SH3 domain is essential for PLC-γ1-mediated cellular signaling, including its own catalytic activity by protein-protein interaction.

Original languageEnglish
Pages (from-to)259-265
Number of pages7
JournalMolecules and cells
Volume6
Issue number3
Publication statusPublished - 1996 Jun 30

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NIH 3T3 Cells
Inositol Phosphates
src Homology Domains
Type C Phospholipases
Tyrosine
Proteins
Cell Shape
Isoenzymes
Carrier Proteins
Complementary DNA
Phosphorylation
Serum

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Chang, Jong Soo ; Min, Do Sik ; Bae, Sun Sik ; Kim, Jae Ho ; Lee, Young Han ; Ryu, Sung Ho ; Suh, Pann Ghill. / Overexpression of SH2-SH2-SH3 domain of phospholipase C-γ1 blocks PDGF-induced inositol phosphate generation in NIH 3T3 cells. In: Molecules and cells. 1996 ; Vol. 6, No. 3. pp. 259-265.
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abstract = "Src homology (SH) 2 and 3 domains are known to be binding motifs for protein-protein interaction in signaling molecules. Among several PLC isozymes, only PLC-γ contains the SH domain between the X and Y domains, which are known to have catalytic activity. To elucidate the functional roles of the SH2-SH2-SH3 domain of PLC-γ1 in cellular signaling, we constructed a truncated cDNA encoding the SH2-SH2-SH3 domain of PLC-γ1 (p60SH2/SH3) and expressed it in NIH 3T3 cells. Cells expressing p60SH2/SH3 did not show any change in cell shape no oncogenesity. Even though in a serum depleted condition, a portion of p60SH2/SH3 existed as constitutively phosphorylated on its tyrosine residues. Furthermore, cells expressing p60SH2/SH3 did not respond to PDGF-induced IPs formation whereas vector transfected control cells showed dose-dependent IPs generation upon PDGF stimulation. The tyrosine phosphorylation level of endogenous PLC-γ1 by PDGF, however, was comparable to that of the control cells. On the other hand, IPs accumulation by PLC-β activation occurred to a comparable level. Taken together, p60SH2/SH3 molecules selectively inhibited the IPs accumulation catalyzed by PLC-γ1. This result suggests that the SH2-SH2-SH3 domain is essential for PLC-γ1-mediated cellular signaling, including its own catalytic activity by protein-protein interaction.",
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Overexpression of SH2-SH2-SH3 domain of phospholipase C-γ1 blocks PDGF-induced inositol phosphate generation in NIH 3T3 cells. / Chang, Jong Soo; Min, Do Sik; Bae, Sun Sik; Kim, Jae Ho; Lee, Young Han; Ryu, Sung Ho; Suh, Pann Ghill.

In: Molecules and cells, Vol. 6, No. 3, 30.06.1996, p. 259-265.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Overexpression of SH2-SH2-SH3 domain of phospholipase C-γ1 blocks PDGF-induced inositol phosphate generation in NIH 3T3 cells

AU - Chang, Jong Soo

AU - Min, Do Sik

AU - Bae, Sun Sik

AU - Kim, Jae Ho

AU - Lee, Young Han

AU - Ryu, Sung Ho

AU - Suh, Pann Ghill

PY - 1996/6/30

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N2 - Src homology (SH) 2 and 3 domains are known to be binding motifs for protein-protein interaction in signaling molecules. Among several PLC isozymes, only PLC-γ contains the SH domain between the X and Y domains, which are known to have catalytic activity. To elucidate the functional roles of the SH2-SH2-SH3 domain of PLC-γ1 in cellular signaling, we constructed a truncated cDNA encoding the SH2-SH2-SH3 domain of PLC-γ1 (p60SH2/SH3) and expressed it in NIH 3T3 cells. Cells expressing p60SH2/SH3 did not show any change in cell shape no oncogenesity. Even though in a serum depleted condition, a portion of p60SH2/SH3 existed as constitutively phosphorylated on its tyrosine residues. Furthermore, cells expressing p60SH2/SH3 did not respond to PDGF-induced IPs formation whereas vector transfected control cells showed dose-dependent IPs generation upon PDGF stimulation. The tyrosine phosphorylation level of endogenous PLC-γ1 by PDGF, however, was comparable to that of the control cells. On the other hand, IPs accumulation by PLC-β activation occurred to a comparable level. Taken together, p60SH2/SH3 molecules selectively inhibited the IPs accumulation catalyzed by PLC-γ1. This result suggests that the SH2-SH2-SH3 domain is essential for PLC-γ1-mediated cellular signaling, including its own catalytic activity by protein-protein interaction.

AB - Src homology (SH) 2 and 3 domains are known to be binding motifs for protein-protein interaction in signaling molecules. Among several PLC isozymes, only PLC-γ contains the SH domain between the X and Y domains, which are known to have catalytic activity. To elucidate the functional roles of the SH2-SH2-SH3 domain of PLC-γ1 in cellular signaling, we constructed a truncated cDNA encoding the SH2-SH2-SH3 domain of PLC-γ1 (p60SH2/SH3) and expressed it in NIH 3T3 cells. Cells expressing p60SH2/SH3 did not show any change in cell shape no oncogenesity. Even though in a serum depleted condition, a portion of p60SH2/SH3 existed as constitutively phosphorylated on its tyrosine residues. Furthermore, cells expressing p60SH2/SH3 did not respond to PDGF-induced IPs formation whereas vector transfected control cells showed dose-dependent IPs generation upon PDGF stimulation. The tyrosine phosphorylation level of endogenous PLC-γ1 by PDGF, however, was comparable to that of the control cells. On the other hand, IPs accumulation by PLC-β activation occurred to a comparable level. Taken together, p60SH2/SH3 molecules selectively inhibited the IPs accumulation catalyzed by PLC-γ1. This result suggests that the SH2-SH2-SH3 domain is essential for PLC-γ1-mediated cellular signaling, including its own catalytic activity by protein-protein interaction.

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