Tissue-specific distribution and subcellular distribution of phospholipase D in rat: evidence for distinct RhoA- and ADP-ribosylation factor (ARF)-regulated isoenzymes

Provost, Joseph; Fudge, J. F.; Israelit, Sallie; Siddiqi, Abdur Rehman; Exton, John H.

doi:10.1042/bj3190285

Cited by 78 publications

(53 citation statements)

References 52 publications

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“…This is based on differences in substrate specificity; in the regulation of the enzyme from different tissues and subcellular fractions by PKC, PIP 2 , fatty acids, Rho, and ARF; and in the effects of divalent cations, detergents, and pH (1,30,31). Although it is probable that the regulation of PLD is complex, the results strongly suggest the presence of several isozymes.…”

Section: And References Therein)mentioning

confidence: 68%

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Cloning and Characterization of Phospholipase D from Rat Brain

Park

Provost

Bae

et al. 1997

Journal of Biological Chemistry

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141

109

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In membranes from COS-7 cells expressing rPLD, addition of myristoylated ADP-ribosylation factor (ARF) and RhoA in vitro stimulated PLD activity. The effect of ARF was greater than that of RhoA, although the concentrations for half-maximal stimulation (0.08 -0.2 M) were similar. Membranes isolated from cells expressing rPLD plus L71ARF3 and/or V14RhoA also showed higher PLD activity but no synergism between the two G proteins. Addition of phorbol ester and protein kinase C ␣ (PKC␣) also stimulated PLD activity in membranes from COS-7 cells expressing rPLD, but it had no effect on the activity in control (vector) membranes and did not enhance the effects of constitutively active ARF or Rho. The stimulation by PKC␣ did not require ATP and was not increased by addition of this nucleotide. No synergism between ARF and Rho and between these and PKC␣ on PLD activity was observed when these were added to membranes from cells expressing rPLD. Oleate inhibited the PLD activity of membranes from both control and rPLD-expressing cells.In summary, these results indicate that in vitro, rPLD is stimulated by ARF, RhoA, and PKC␣ and inhibited by oleate. However, in intact COS-7 cells, ARF activates endogenous PLD but not rPLD, whereas the reverse is true for RhoA. In addition, the effects of phorbol ester are much greater in the intact cells. It is concluded that the regulation of rPLD in intact COS-7 cells differs significantly from that seen in vitro; possible reasons for this are discussed.

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Section: And References Therein)mentioning

confidence: 68%

“…[2-palmitoyl-9,10-3 H]Phosphatidylcholine and [9, H]myristic acid were purchased from NEN Life Science Products. PKC␣ was purchased from Panvera Corp. Phosphatidylinositol 4,5-bisphosphate (PIP 2 ), RhoA, and recombinant myristoylated ARF3 (mARF) were prepared as described previously (31). SDS-polyacrylamide gels were purchased from Novex.…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of Phospholipase D from Rat Brain

Park

Provost

Bae

et al. 1997

Journal of Biological Chemistry

Self Cite

141

109

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“…Rac1 and Cdc42 are also involved in the activation of PLD in liver and brain (8,9). While the synergistic activation by RhoA and ARF suggests convergence of regulatory mechanisms on one single PLD isoform (8), additional biochemical studies indicate the presence of distinct RhoA-and ARF-regulated PLD isoforms in HL-60 cells and liver (10,11). RhoA-and ARF-responsive PLD activities are observed in the plasma membrane, nuclei, Golgi, and endoplasmic reticulum (10,(12)(13)(14)(15).…”

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confidence: 99%

Human Phospholipase D1 Can Be Tyrosine-phosphorylated in HL-60 Granulocytes

Marcil¹,

Harbour

Naccache

et al. 1997

Journal of Biological Chemistry

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The human phospholipase D1 (hPLD1) has recently been cloned. Although recent data have implicated PLD in receptor-stimulated secretion, the regulation of the activity of PLD enzymes remains to be clarified. Purified hPLD1 is activated by several cytosolic cofactors among which are protein kinase C␣, ARF, and RhoA. In human granulocytes, a strong correlation between tyrosine phosphorylation of proteins and PLD activity has been established. In this study, the presence of hPLD1 in HL-60 granulocytes and its phosphorylation on tyrosine residues have been studied. We generated antipeptide antibodies (Abs) specific for hPLD1 but not PLD2 as shown by Western blotting (WB) of recombinant PLD1 and PLD2. These Abs identified the presence of hPLD1 in HL-60 cells with the bulk of it being detected in the membranes and only a minor fraction in the cytosol. The hPLD1 Abs detected a major band at 120 kDa (PLD1a) and a minor band at 115 kDa (PLD1b). The specificity of the Abs was confirmed using PLD antisera neutralized with the immunizing peptides. The two forms of hPLD1 were consistently detected by immunoprecipitation under nondenaturing and denaturing conditions following a WB analysis with hPLD1 Abs. Following exposure of HL-60 cells to peroxides of vanadate (V 4؉ -OOH), an inhibitor of tyrosine phosphatases, hPLD1 was immunoprecipitated under nondenaturing conditions from HL-60 cell lysates and assayed for tyrosine phosphorylation by WB. hPLD1 comigrated with a 120-kDa tyrosine phosphorylated protein by gel electrophoresis. Other tyrosine-phosphorylated peptides of 160, 140, 135, 90, and 75-80 kDa were also detected in hPLD1 immune complexes. hPLD1 and the associated tyrosine-phosphorylated proteins were not immunoprecipitated by neutralized hPLD1 Abs. Using denaturing conditions, the PLD immunoprecipitates were sequentially immunoblotted with anti-PLD1 and anti-phosphotyrosine Abs. PLD1a and PLD1b were detected, and the major PLD1a protein was superimposable with a major tyrosine-phosphorylated protein detected at 120 kDa. Conversely, PLD1a and PLD1b were recovered, at least in part, in the anti-phosphotyrosine immunoprecipitates. These results provide evidence that two PLD1 forms are expressed in human granulocytes. Furthermore, in response to stimulation by V 4؉ -OOH, PLD1 was tyrosinephosphorylated and associated with several, presently undefined, tyrosine-phosphorylated proteins.

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“…Therefore, learning more about the localization of the enzyme and its product PA is important to the understanding of its function. PLD activities have been reported for diverse subcellular organelles including plasma membrane, Golgi vesicles, endoplasmic reticulum, secretory vesicles, and the nuclear membrane (15)(16)(17)(18)(19)(20). There are consequently the following questions to be answered: which isozyme of PLD localizes to a certain organelle, how is the isozyme differentially regulated, and what is its role?…”

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confidence: 99%