scholarly journals Plastid Deficient 1 Is Essential for the Accumulation of Plastid-Encoded RNA Polymerase Core Subunit β and Chloroplast Development in Arabidopsis

2021 ◽  
Vol 22 (24) ◽  
pp. 13648
Author(s):  
Zhipan Yang ◽  
Mingxin Liu ◽  
Shunhua Ding ◽  
Yi Zhang ◽  
Huixia Yang ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Chloroplast Development ◽  
Direct Interaction ◽  
Immunoblot Analysis ◽  
Regulation Mechanism ◽  
Plastid Development ◽  
Essential Components ◽  
Complex Regulation ◽  
Identification And Characterization

Plastid-encoded RNA polymerase (PEP)-dependent transcription is an essential process for chloroplast development and plant growth. It is a complex event that is regulated by numerous nuclear-encoded proteins. In order to elucidate the complex regulation mechanism of PEP activity, identification and characterization of PEP activity regulation factors are needed. Here, we characterize Plastid Deficient 1 (PD1) as a novel regulator for PEP-dependent gene expression and chloroplast development in Arabidopsis. The PD1 gene encodes a protein that is conserved in photoautotrophic organisms. The Arabidopsis pd1 mutant showed albino and seedling-lethal phenotypes. The plastid development in the pd1 mutant was arrested. The PD1 protein localized in the chloroplasts, and it colocalized with nucleoid protein TRXz. RT-quantitative real-time PCR, northern blot, and run-on analyses indicated that the PEP-dependent transcription in the pd1 mutant was dramatically impaired, whereas the nuclear-encoded RNA polymerase-dependent transcription was up-regulated. The yeast two-hybrid assays and coimmunoprecipitation experiments showed that the PD1 protein interacts with PEP core subunit β (PEP-β), which has been verified to be essential for chloroplast development. The immunoblot analysis indicated that the accumulation of PEP-β was barely detected in the pd1 mutant, whereas the accumulation of the other essential components of the PEP complex, such as core subunits α and β′, were not affected in the pd1 mutant. These observations suggested that the PD1 protein is essential for the accumulation of PEP-β and chloroplast development in Arabidopsis, potentially by direct interaction with PEP-β.

scholarly journals Identification and Characterization of the Direct Interaction between Methotrexate (MTX) and High-Mobility Group Box 1 (HMGB1) Protein

PLoS ONE ◽  
2013 ◽  
Vol 8 (5) ◽  
pp. e63073 ◽  
Author(s):  
Yuki Kuroiwa ◽  
Yoichi Takakusagi ◽  
Tomoe Kusayanagi ◽  
Kouji Kuramochi ◽  
Takahiko Imai ◽  
...  
Keyword(s):  
High Mobility ◽  
Direct Interaction ◽  
High Mobility Group ◽  
Hmgb1 Protein ◽  
Identification And Characterization

Identification and characterization of a fibronectin-binding protein from Clostridium difficile

Microbiology ◽  
2003 ◽  
Vol 149 (10) ◽  
pp. 2779-2787 ◽  
Author(s):  
Claire Hennequin ◽  
Claire Janoir ◽  
Marie-Claude Barc ◽  
Anne Collignon ◽  
Tuomo Karjalainen
Keyword(s):  
Clostridium Difficile ◽  
Binding Protein ◽  
Immunoblot Analysis ◽  
Vero Cells ◽  
Cell Fractionation ◽  
Fibronectin Binding Protein ◽  
Fibronectin Binding ◽  
The One ◽  
Identification And Characterization

A 68 kDa fibronectin-binding protein (Fbp68) from Clostridium difficile displaying significant homology to several established or putative Fbps from other bacteria was identified. The one-copy gene is highly conserved in C. difficile isolates. Fbp68 was expressed in Escherichia coli in fusion with glutathione S-transferase; the fusion protein and the native Fbp68 were purified. Immunoblot analysis and cell fractionation experiments revealed that Fbp68 is present on the surface of the bacteria. Far-immuno dot-blotting demonstrated that Fbp68 was capable of fixing fibronectin. Indirect immunofluorescence and ELISA were employed to demonstrate that C. difficile could bind both soluble and immobilized fibronectin. With competitive adherence inhibition assays it was shown that antibodies raised against Fbp68 partially inhibited attachment of C. difficile to fibronectin and Vero cells. Furthermore, Vero cells could fix purified membrane-immobilized Fbp68. Thus Fbp68 appears to be one of the several adhesins identified to date in C. difficile.

Identification and characterization of Dicer-like, Argonaute and RNA-dependent RNA polymerase gene families in maize

2011 ◽  
Vol 30 (7) ◽  
pp. 1347-1363 ◽  
Author(s):  
Yexiong Qian ◽  
Ying Cheng ◽  
Xiao Cheng ◽  
Haiyang Jiang ◽  
Suwen Zhu ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Gene Families ◽  
Polymerase Gene ◽  
Rna Dependent Rna Polymerase ◽  
Identification And Characterization ◽  
Rna Polymerase Gene

scholarly journals Identification and characterization of a novel spore-associated subtilase from Thermoactinomyces sp. CDF

Microbiology ◽  
2009 ◽  
Vol 155 (11) ◽  
pp. 3661-3672 ◽  
Author(s):  
Guyue Cheng ◽  
Peiwei Zhao ◽  
Xiao-Feng Tang ◽  
Bing Tang
Keyword(s):  
Inclusion Bodies ◽  
Immunoblot Analysis ◽  
Gene Encoding ◽  
Ph Values ◽  
Mature Enzyme ◽  
Ph Conditions ◽  
Coat Layer ◽  
Identification And Characterization ◽  
Mature Protease

A gene encoding a spore-associated subtilase, designated protease CDF, was cloned from Thermoactinomyces sp. CDF and expressed in Escherichia coli. The enzyme gene is translated as a proform consisting of a 94 aa propeptide and a 283 aa mature protease domain. Phylogenetic analysis revealed that this enzyme belonged to the subtilisin family, but could not be grouped into any of its six known subfamilies. The mature protease CDF has an unusually high content of charged residues, which are mainly distributed on the enzyme surface. The recombinant proform of protease CDF formed inclusion bodies, but could be efficiently converted to the mature enzyme when the inclusion bodies were dissolved in alkaline buffers. The proform underwent a two-step maturation process, wherein the N-terminal part (85 residues) of the propeptide was autoprocessed intramolecularly, and the remaining 9-residue peptide was further processed intermolecularly. Protease CDF exhibited optimal proteolytic activity at 50–55 °C and pH 10.5–11.0. The enzyme was stable under high-pH conditions (pH 11.0–12.0), and NaCl could stabilize the enzyme at lower pH values. In addition, the enzyme was not dependent on calcium for either maturation or stability. By immunoblot analysis, protease CDF was found to be associated with spores, and could be extracted from the spores with 2 M KCl and alkaline buffers without damaging the coat layer, demonstrating that the protease CDF is located on the surface of the spore coat.

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