scholarly journals Topography of transcription: path of the leading end of nascent RNA through the Escherichia coli transcription complex.

1983 ◽  
Vol 80 (14) ◽  
pp. 4238-4242 ◽  
Author(s):  
M. M. Hanna ◽  
C. F. Meares
Keyword(s):  
Escherichia Coli ◽  
Nascent Rna

scholarly journals Chemical roadblocking of DNA transcription for nascent RNA display

2020 ◽  
Vol 295 (19) ◽  
pp. 6401-6412
Author(s):  
Eric J. Strobel ◽  
John T. Lis ◽  
Julius B. Lucks
Keyword(s):  
Escherichia Coli ◽  
Rna Structure ◽  
Triethylene Glycol ◽  
Site Specific ◽  
Rna Molecules ◽  
Protein Factor ◽  
Nascent Rna ◽  
Dna Strand ◽  
And Function

Site-specific arrest of RNA polymerases (RNAPs) is fundamental to several technologies that assess RNA structure and function. Current in vitro transcription “roadblocking” approaches inhibit transcription elongation by blocking RNAP with a protein bound to the DNA template. One limitation of protein-mediated transcription roadblocking is that it requires inclusion of a protein factor extrinsic to the minimal in vitro transcription reaction. In this work, we developed a chemical approach for halting transcription by Escherichia coli RNAP. We first established a sequence-independent method for site-specific incorporation of chemical lesions into dsDNA templates by sequential PCR and translesion synthesis. We then show that interrupting the transcribed DNA strand with an internal desthiobiotin-triethylene glycol modification or 1,N6-etheno-2′-deoxyadenosine base efficiently and stably halts Escherichia coli RNAP transcription. By encoding an intrinsic stall site within the template DNA, our chemical transcription roadblocking approach enables display of nascent RNA molecules from RNAP in a minimal in vitro transcription reaction.

scholarly journals The interaction between  70 and the  -flap of Escherichia coli RNA polymerase inhibits extension of nascent RNA during early elongation

2005 ◽  
Vol 102 (12) ◽  
pp. 4488-4493 ◽  
Author(s):  
B. E. Nickels ◽  
S. J. Garrity ◽  
V. Mekler ◽  
L. Minakhin ◽  
K. Severinov ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Nascent Rna
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