PU.1 Is Dominant and HAF-1 Supplementary for Activation of the gp91phox Promoter in Human Monocytic PLB-985 Cells.

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概要

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• Gp91phox is a key component of the phagocyte NADPH oxidase. Mutations of its promoter found in patients with chronic granulomatous disease cause deficient binding of PU. 1 and HAF-1. Because the two factors bind to the same site (Pu box) of the promoter, we attempted to clarify their relative in vivo contributions to activation of the gp91phox promoter in monocytically differentiated PLB-985 cells using a dual luciferase reporter assay and a gel shift competition assay. We found that the activity of a series of singlepoint-mutated promoters increases or decreases according to an increase or decrease, respectively, in the affinity of the promoters to PU. 1 but not to HAF-1. Two of 7 mutants showing weak binding affinity to PU.l exhibited moderate promoter activity and normal binding affinity for HAF-1. These results suggest PU. 1 is the dominant activator and HAF-1 is supplementary. The increased promoter activity of single-, double-, and triplepoint-mutated constructs with sequences closer to that of the Ets-binding element correlates with their binding affinity to PU. 1 but not to HAF-1, supporting that PU. 1 is a more efficient activator than HAF-1. In contrast to co-expressed wild-type PU. 1, dominantnegative PU. 1 significantly inhibited the activity of a PU. 1-optimised gp91phox promoter construct. Therefore, we conclude that PU. 1 and HAF-1 binding to the Pu box is dominant and supplementary, respectively, for activation of the gp91phox promoter in human monocytic cells.

• 社団法人 日本生化学会の論文

著者

• SUZUKI Shoichi

  Department of Host-Defense Biochemistry, Institute of Tropical Medicine, Nagasaki University

• Hao Li

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• SUZUKI Norio

  Pharmaceutical Formulation Research Laboratory, Daiichi Pharmaceutical Co., Ltd.

• KIKUCHI Hiroshi

  Pharmaceutical Formulation Research Laboratory, Daiichi Pharmaceutical Co., Ltd.

• Suzuki Shoichi

  Department Of Agricultural Science Ishikawa Agricultural College

• Nakamura Michio

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• Fan Chun

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• Yang Dan

  Department Of Chemistry The University Of Hong Kong

• Fujii Yoshito

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• Islam Rafiqul

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• Rusvai Erzsebet

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

• Kumatori Atsushi

  Department Of Host-defense Biochemistry Institute Of Tropical Medicine Nagasaki University

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