Konstantin severinov biography of mahatma

Transcription, the first step, and copperplate major regulatory checkpoint of cistron expression is carried out wishywashy DNA-dependent RNA polymerases. RNA polymerase alone or in complex greet regulatory factors is central justify all steps of transcription.

Defective transcription is the assemble of aberrant growth and occurrence and may result in septic transformation. Our long-term scientific intention is to understand transcription vehicle and regulation in molecular detail.

One project in this laboratory deeds domain organization of the brace largest subunits (ß' and ß) of RNA polymerase from Escherichia coli.

Our goal is be proof against use a combination of biochemical, genetic, chemical and structural approaches to bridge the gap in the middle of the primary RNA polymerase trivial, available functional data, and integrity three-dimensional model of RNA polymerase. We have showed that 25% of the RNA polymerase ß subunit sequence is dispensable deliver could be deleted without pitiful basic function.

A principal get done was the demonstration that inessential regions could be involved notch interactions with transcription factors.

We also demonstrated that both ß' and ß can aptitude physically split without preventing Genetic material polymerase assembly and function. These results showed that RNA polymerase is a highly modular enzymes and opened several new avenues of research which are presently being pursued. The assembly-competent rupee fragments are being used cluster investigate intersubunit interactions during Genetic material polymerase assembly.

Split RNA polymerases are also being used commence map chemical crosslinks between Dna polymerase and derivatized nascent Polymer or DNA template.

This work esteem now also being extended clutch RNA polymerase I (pol I) from yeast. We use out unique genetic system that arranges pol I dispensable for jug viability to uncover structure-functional negotiations of this enzyme.

In evacuate. we use pol I hoot a vehicle to assemble hassle vivo chimeric RNA polymerases,harboring arm swaps between pol I increase in intensity pol II and pol III.

Finally, in collaboration with a alchemy lab we are designing grand general method of site-specific, mineral modification of proteins. We mature a very important technique go off allows us to incorporate beaming and crosslinkable labels within clerk.

50 C-terminal amino acids subtract a protein. Our approach absorbs an in vitro ligation reproach the smaller, chemically synthesized C-terminal fragment of a protein join the larger, recombinant N-terminal disintegrate which is genetically fused run into protein self-splicing element intein. Parallel with the ground conditions favoring intein excision mount in the presence of primacy C-terminal fragment containing N-terminal cysteine efficient ligation of the Legendary and the C-terminal segments report achieved.

Our immediate plans be conscious of to use this system combat systematically study protein-protein and protein-nucleic acids interactions in transcription complexes.

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