Increased Activity of Small GTP-binding Protein-dependent Phospholipase D during Differentiation in Human Promyelocytic Leukemic HL60 Cells*
Increased Activity of Small GTP-binding Protein-dependent Phospholipase D during Differentiation in Human Promyelocytic Leukemic HL60 Cells*
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人早幼粒细胞白血病 HL60 细胞分化过程中小 GTP 结合蛋白依赖性磷脂酶 D 的活性增加*
DOI：
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发表时间：
1997
影响因子：
4.8
通讯作者：
Y. Nozawa
中科院分区：
文献类型：
--
作者：
K. Ohguchi;S. Nakashima;Z. Tan;Y. Banno;Shuji Dohi;Y. Nozawa
二丁酰环腺苷酸（dbcAMP）和全反式维甲酸诱导人早幼粒细胞白血病HL 60细胞分化为粒细胞样细胞。在细胞裂解液和体外重组体系中，dbcAMP或全反式维甲酸处理可上调鸟苷5′-O-（3-硫代三磷酸）（GTPγS）诱导的磷脂酶D（PLD）活性。在本研究中，研究了分化过程中PLD活性增加的机制。Western blot分析显示，ADP-核糖基化因子，Rac 2，Cdc 42 Hs，但不是RhoA和Rac 1的胞质组分的含量在分化过程中升高。然而，来自未分化细胞的胞质组分在刺激膜PLD活性的能力方面几乎与来自分化细胞的胞质组分同样有效。结果表明，分化细胞膜中依赖GTPγ S的PLD活性明显高于未分化细胞膜，提示分化过程中PLD活性的增加是由于某些膜成分的改变所致。肉毒梭菌ADP-核糖基转移酶C3和C.分别已知为RhoA和Rho家族蛋白抑制剂的艰难梭菌毒素B有效地抑制来自分化细胞的膜中的PLD活性。事实上，膜相关RhoA的量在分化过程中增加。此外，在存在重组ADP-核糖基化因子1的情况下，从分化细胞膜部分纯化的GTPγ S依赖性PLD活性的程度大于从未分化细胞膜部分纯化的GTPγ S依赖性PLD活性的程度。PLD（hPLD 1）mRNA水平被观察到在分化过程中上调，推断通过逆转录-聚合酶链反应。上述结果提示，HL 60细胞粒系分化过程中膜Rho蛋白的增加和PLD自身水平的变化可能是导致依赖GTPγ S的PLD活性增加的部分原因。
In response to dibutyryl cyclic AMP (dbcAMP) and all-trans retinoic acid, human promyelocytic leukemic HL60 cells differentiate into granulocyte-like cells. In cell lysate and in vitro reconstitution system, phospholipase D (PLD) activity in response to guanosine 5′-O-(3-thiotriphosphate) (GTPγS) was up-regulated by dbcAMP or all-trans retinoic acid treatment. In the present study, the mechanism(s) for increased PLD activity during differentiation was examined. Western blot analysis revealed that the contents of ADP-ribosylation factor, Rac2, and Cdc42Hs but not RhoA and Rac1 in the cytosolic fraction were elevated during differentiation. However, the cytosolic fraction from undifferentiated cells was almost equally potent as the cytosolic fraction from differentiated cells in the ability to stimulate membrane PLD activity. It was shown that the GTPγS-dependent PLD activity in membranes from differentiated cells was much higher than that in membranes from undifferentiated cells, suggesting that the increased PLD activity during differentiation was due to alterations in some membrane component(s). Clostridium botulinum ADP-ribosyltransferase C3 and C. difficile toxin B, which are known as inhibitors of RhoA and Rho family proteins, respectively, effectively suppressed PLD activity in membranes from differentiated cells. In fact, the amount of membrane-associated RhoA was increased during differentiation. Furthermore, the extent of GTPγS-dependent PLD activity partially purified from membranes from differentiated cells was greater than that from membranes from undifferentiated cells in the presence of recombinant ADP-ribosylation factor 1. The PLD (hPLD1) mRNA level was observed to be up-regulated during differentiation, as inferred by reverse transcription-polymerase chain reaction. Our results suggest the possibility that the increased Rho proteins in membranes and the changed level of PLD itself may be, at least in part, responsible for the increase in GTPγS-dependent PLD activity during granulocytic differentiation of HL60 cells.
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DOI：
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发表时间：
1992
期刊：
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影响因子：
--
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DOI：
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发表时间：
1992
期刊：
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DOI：
10.1016/s0021-9258(19)47364-x
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1991-09
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DOI：
10.1172/jci115303
发表时间：
1991
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Xie,MS;Jacobs,LS;Dubyak,GR
通讯作者：
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DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Xing,M;Wilkins,PL;McConnell,BK;Mattera,R
通讯作者：
Mattera,R