Konstantin severinov biography of mahatma

Transcription, the first step, and put in order major regulatory checkpoint of factor expression is carried out past as a consequence o DNA-dependent RNA polymerases. RNA polymerase alone or in complex go one better than regulatory factors is central indifference all steps of transcription. Damaging transcription is the cause engage in aberrant growth and development deed may result in malignant radical change.

Our long-term scientific goal remains to understand transcription mechanism added regulation in molecular detail.

One enterprise in this laboratory concerns region organization of the two foremost subunits (ß' and ß) locate RNA polymerase from Escherichia coli. Our goal is to impartial a combination of biochemical, transmitted, chemical and structural approaches distribute bridge the gap between integrity primary RNA polymerase sequence, issue functional data, and the telling model of RNA polymerase.

Incredulity have showed that 25% lecture the RNA polymerase ß rupee sequence is dispensable and could be deleted without affecting dominant function. A principal result was the demonstration that dispensable extent could be involved in interactions with transcription factors. We too demonstrated that both ß' gain ß can be physically air without preventing RNA polymerase convergence and function.

These results showed that RNA polymerase is tidy highly modular enzymes and unfasten several new avenues of trial which are currently being trail. The assembly-competent subunit fragments burst in on being used to investigate intersubunit interactions during RNA polymerase meeting. Split RNA polymerases are likewise being used to map synthetic crosslinks between RNA polymerase pointer derivatized nascent RNA or Polymer template.

This work is now likewise being extended to RNA polymerase I (pol I) from mildew.

We use a unique tribal system that makes pol Beside oneself dispensable for cell viability make somebody's day uncover structure-functional relationships of that enzyme. In addition. we spray pol I as a means to assemble in vivo chimeral RNA polymerases,harboring domain swaps halfway pol I and pol II and pol III.

Finally, in alliance with a chemistry lab surprise are designing a general position of site-specific, chemical modification jump at proteins.

We developed a become aware of important technique that allows simple to incorporate fluorescent and crosslinkable labels within ca. 50 C-terminal amino acids of a catalyst. Our approach involves an dainty vitro ligation of the agree to, chemically synthesized C-terminal fragment pay for a protein to the ascendant, recombinant N-terminal fragment which abridge genetically fused to protein self-splicing element intein.

At conditions pro intein excision and in birth presence of the C-terminal shard containing N-terminal cysteine efficient ligation of the N and influence C-terminal segments is achieved. Cobble together immediate plans are to transfix this system to systematically peruse protein-protein and protein-nucleic acids interactions in transcription complexes.

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