MOLECULAR BIOLOGY ON CYANOBACTERIAL CO_2 PUMPS

蓝藻CO_2泵的分子生物学

• 批准号: 12440228
• 负责人: OGAWA Teruo
• 金额: $ 7.04万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12440228/
• 关键词: CO_2-concentrating mechanism; CO_2 pump; NADPH dehydrogenase; Bicarbonate transporter; SbtA-dependent type; Cmp-dependent type; NdhD3-dependent type; NdhD4-dependent type; ndh遺伝子; NAD(P)Hデヒドロゲナーゼ; らん藻; sbtA; HCO_3^-輸送; cmpA; cbbR; 変異株; cmp

We have identified two CO_2-uptake systems, one constitutive dependent on NdhD3/NdhF3/CupA(Sll 1734) and one low-CO_2-inducible dependent on NdhD4/NdhF4/CupB(Slr 1302), and two types of HCO_3^- transporters, an ABC-type transporter encoded by cmp operon and a novel sodium-dependent transporter encoded by slr 1512 (sbtA) that plays a central role in HCO_3^- uptake in the cyanobacterium Synechocystis sp, strain PCC 6803. Mutants impaired for one of these 4 inorganic carbon-acquisition systems did not show mutant phenotype. Mutants inactivated for both CO_2-uptake systems were unable to grow at pH 7.0 in air although they grew normally at pH 9.0 in air. Additional inactivation of the SbtA-type HCO_3^- transporter abolished the growth at pH 9.0 in air. Using the antibody raised against SbtA, we have shown that SbtA is localized in the cytoplasmic membrane. Immunoprecipitation using this antibody revealed that a protein of about 10 kDa was coprecipitated with SbtA. Amino-acid sequencing revealed that this protein the product of slr 1513 (named as sbtB) cotranscribed with sbtA.

我们鉴定了两个CO_2吸收系统，一个依赖于NdhD 3/NdhF 3/CupA（Sll 1734）和一个依赖于NdhD 4/NdhF 4/CupB的低CO_2诱导型（Slr 1302），以及两种类型的HCO_3^-转运蛋白，一种由CMP操纵子编码的ABC型转运蛋白和一种由slr 1512（sbtA）编码的新型钠依赖转运蛋白，其在HCO_3^-在蓝细菌集胞藻属（Synechocystis sp）菌株PCC 6803中的吸收。这4个无机碳获取系统之一受损的突变体没有表现出突变表型。两种CO_2吸收系统失活的突变体在pH9.0的空气中生长正常，但在pH7.0的空气中不能生长。在pH9.0的空气中，进一步灭活SbtA型HCO_3^-转运蛋白可抑制其生长。使用针对SbtA的抗体，我们已经表明SbtA定位于细胞质膜中。使用该抗体的免疫沉淀显示，约10 kDa的蛋白质与SbtA共沉淀。氨基酸序列分析表明，该蛋白是slr 1513的产物（命名为sbtB），与sbtA共转录。

项目成果

Katoh,H.,Hagino,N.,Grossman,A.R.,and Ogawa,T.: "Genes essential to iron transport in the cyanobacterium Synechocystis sp. Strain PCC6803"Journal of Bacteriology. 186(印刷中). (2000)

Katoh, H.、Hagino, N.、Grossman, A.R. 和 Okawa, T.：“蓝细菌集胞藻菌株 PCC6803 中铁转运所必需的基因”《细菌学杂志》186（印刷中）。

Ohkawa, H., Sonoda, M., Shibata, M., Ogawa, T.: "Localization of NAD(P)H dehydrogenase in the cyanobacterium Synechocystis sp. PCC 6803"J. Bacteriol.. 183. 4938-4939 (2001)

Ohkawa, H.、Sonoda, M.、Shibata, M.、Okawa, T.：“NAD(P)H 脱氢酶在蓝藻集胞藻 PCC 6803 中的定位”J。

Ohkawa, H., Pakrasi, H.B.and Ogawa, T.: "Two types of functionally distinct NAD(P)H dehydrogenases in Synechocystis sp.strain PCC6803"J.Biol.Chem. 275. 31630-31634 (2000)

Ohkawa, H.、Pakrasi, H.B. 和 Okawa, T.：“集胞藻菌株 PCC6803 中两种功能不同的 NAD(P)H 脱氢酶”J.Biol.Chem。

Shibata, M., Ohkawa, H., Kaneko, T., Fukuzawa, H., Tabata, S., Kaplan, A., Ogawa, T.: "Distinct constitutive and low CO_2-induced CO_2 uptake systems in cyanobacteria: Genes involved and their phylogenetic relationship with homologous genes in other organ

Shibata, M.、Ohkawa, H.、Kaneko, T.、Fukuzawa, H.、Tabata, S.、Kaplan, A.、Okawa, T.：“蓝藻中独特的组成型和低 CO_2 诱导的 CO_2 吸收系统：基因

Omata, T., Gohta, S., Takahashi, Y., Harano, Y., Maeda, S.: "Involvement of a CbbR homolog in low CO2 induced activation of the bicarbonate transporter operon in cyanobacteria"J.Bacteriol.. 183. 1891-1898 (2001)

Omata，T.，Gohta，S.，Takahashi，Y.，Harano，Y.，Maeda，S.：“CbbR 同源物参与低 CO2 诱导蓝藻中碳酸氢盐转运蛋白操纵子的激活”J.Bacteriol.. 183