scholarly journals In vitro assembly of an archaeal D-L-N RNA polymerase subunit complex reveals a eukaryote-like structural arrangement

1998 ◽  
Vol 26 (24) ◽  
pp. 5562-5567 ◽  
Author(s):  
J. Eloranta
Keyword(s):  
Rna Polymerase ◽  
Structural Arrangement ◽  
In Vitro Assembly

scholarly journals The RNA polymerase factory: a robotic in vitro assembly platform for high-throughput production of recombinant protein complexes

2007 ◽  
Vol 36 (1) ◽  
pp. 245-252 ◽  
Author(s):  
Sven Nottebaum ◽  
Lin Tan ◽  
Dominika Trzaska ◽  
Hannah C. Carney ◽  
Robert O. J. Weinzierl
Keyword(s):  
Recombinant Protein ◽  
Rna Polymerase ◽  
High Throughput ◽  
Protein Complexes ◽  
In Vitro Assembly

scholarly journals In vitro assembly and proteomic analysis of RNA polymerase II complexes

Methods ◽  
2019 ◽  
Vol 159-160 ◽  
pp. 96-104 ◽  
Author(s):  
Yoo Jin Joo ◽  
Scott B. Ficarro ◽  
Jarrod A. Marto ◽  
Stephen Buratowski
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Proteomic Analysis ◽  
In Vitro Assembly

scholarly journals In Vitro Assembly of PB2 with a PB1-PA Dimer Supports a New Model of Assembly of Influenza A Virus Polymerase Subunits into a Functional Trimeric Complex

2005 ◽  
Vol 79 (13) ◽  
pp. 8669-8674 ◽  
Author(s):  
Tao Deng ◽  
Jane Sharps ◽  
Ervin Fodor ◽  
George G. Brownlee
Keyword(s):  
Rna Polymerase ◽  
Influenza A ◽  
Viral Rna ◽  
Trimeric Complex ◽  
Virus Rna ◽  
Proposed Model ◽  
In Vitro Assembly ◽  
Rna Promoter ◽  
Sequential Assembly

ABSTRACT Influenza virus RNA-dependent RNA polymerase is a heterotrimeric complex of PB1, PB2, and PA. We show that the individually expressed PB2 subunit can be assembled with the coexpressed PB1-PA dimer in vitro into a transcriptionally active complex. Furthermore, we demonstrate that a model viral RNA promoter can bind to the PB1-PA dimer prior to assembly with PB2. Our results are consistent with a recently proposed model for the sequential assembly of viral RNA polymerase complex in which the PB1-PA dimeric complex and the PB2 monomer are transported into the nucleus separately and then assembled in the nucleus.

In vitro assembly of 2-D crystals from partially purified EA1 protein secreted by Bacillus anthracis strain Delta Sterne-1

1994 ◽  
Vol 52 ◽  
pp. 276-277
Author(s):  
Mary Beth Downs ◽  
Wilson Ribot ◽  
Joseph W. Farchaus
Keyword(s):  
Cell Wall ◽  
Molecular Sieves ◽  
Cell Envelope ◽  
Single Species ◽  
Supporting Cell ◽  
Surface Layers ◽  
Rabbit Igg ◽  
In Vitro Assembly ◽  
Covalently Linked

Many bacteria possess surface layers (S-layers) that consist of a two-dimensional protein lattice external to the cell envelope. These S-layer arrays are usually composed of a single species of protein or glycoprotein and are not covalently linked to the underlying cell wall. When removed from the cell, S-layer proteins often reassemble into a lattice identical to that found on the cell, even without supporting cell wall fragments. S-layers exist at the interface between the cell and its environment and probably serve as molecular sieves that exclude destructive macromolecules while allowing passage of small nutrients and secreted proteins. Some S-layers are refractory to ingestion by macrophages and, generally, bacteria are more virulent when S-layers are present.When grown in rich medium under aerobic conditions, B. anthracis strain Delta Sterne-1 secretes large amounts of a proteinaceous extractable antigen 1 (EA1) into the growth medium. Immunocytochemistry with rabbit polyclonal anti-EAl antibody made against the secreted protein and gold-conjugated goat anti-rabbit IgG showed that EAI was localized at the cell surface (fig 1), which suggests its role as an S-layer protein.

scholarly journals Functional dissection of the catalytic mechanism of mammalian RNA polymerase II

2005 ◽  
Vol 83 (4) ◽  
pp. 497-504 ◽  
Author(s):  
Benoit Coulombe ◽  
Marie-France Langelier
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Catalytic Mechanism ◽  
Mutational Analysis ◽  
Structural Elements ◽  
Catalytic Mechanisms ◽  
Rnap Ii ◽  
Affinity Peptide

High resolution X-ray crystal structures of multisubunit RNA polymerases (RNAP) have contributed to our understanding of transcriptional mechanisms. They also provided a powerful guide for the design of experiments aimed at further characterizing the molecular stages of the transcription reaction. Our laboratory used tandem-affinity peptide purification in native conditions to isolate human RNAP II variants that had site-specific mutations in structural elements located strategically within the enzyme's catalytic center. Both in vitro and in vivo analyses of these mutants revealed novel features of the catalytic mechanisms involving this enzyme.Key words: RNA polymerase II, transcriptional mechanisms, mutational analysis, mRNA synthesis.

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