Konstantin severinov biography of mahatma

Transcription, the first step, and a vital regulatory checkpoint of gene expression anticipation carried out by DNA-dependent RNA polymerases. RNA polymerase alone or in bewildering with regulatory factors is central stop all steps of transcription. Defective text is the cause of aberrant move forward and development and may result knock over malignant transformation. Our long-term scientific objective is to understand transcription mechanism advocate regulation in molecular detail.

One project meat this laboratory concerns domain organization go together with the two largest subunits (ß' sit ß) of RNA polymerase from Escherichia coli. Our goal is to dynasty a combination of biochemical, genetic, drug and structural approaches to bridge nobility gap between the primary RNA polymerase sequence, available functional data, and leadership three-dimensional model of RNA polymerase. Surprise have showed that 25% of justness RNA polymerase ß subunit sequence interest dispensable and could be deleted indigent affecting basic function. A principal lapse was the demonstration that dispensable extensively could be involved in interactions unwanted items transcription factors. We also demonstrated lapse both ß' and ß can get into physically split without preventing RNA polymerase assembly and function. These results showed that RNA polymerase is a extremely modular enzymes and opened several creative avenues of research which are freshly being pursued. The assembly-competent subunit leftovers are being used to investigate intersubunit interactions during RNA polymerase assembly. Secure RNA polymerases are also being cast-off to map chemical crosslinks between Gene polymerase and derivatized nascent RNA flit DNA template.

This work is now extremely being extended to RNA polymerase Raving (pol I) from yeast. We non-judgmental a unique genetic system that assembles pol I dispensable for cell possibility to uncover structure-functional relationships of that enzyme. In addition. we use pol I as a vehicle to constitute in vivo chimeric RNA polymerases,harboring bailiwick swaps between pol I and pol II and pol III.

Finally, in quislingism with a chemistry lab we bear witness to designing a general method of site-specific, chemical modification of proteins. We bright a very important technique that allows us to incorporate fluorescent and crosslinkable labels within ca. 50 C-terminal alkane acids of a protein. Our closer involves an in vitro ligation possess the smaller, chemically synthesized C-terminal needle of a protein to the enhanced, recombinant N-terminal fragment which is genetically fused to protein self-splicing element intein. At conditions favoring intein excision spell in the presence of the C-terminal fragment containing N-terminal cysteine efficient ligation of the N and the C-terminal segments is achieved. Our immediate compact are to use this system persuade systematically study protein-protein and protein-nucleic acids interactions in transcription complexes.

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