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Structural and functional studies on the negative elongation factor
Structural and functional studies on the negative elongation factor
In many metazoans RNA polymerase II is stably paused after transcribing 20-60 nucleotides of a gene and awaits controlled release. The four subunit Negative Elongation Factor (NELF) is essential to this process which is called promoter-proximal pausing. In this work, a highly conserved NELF subcomplex consisting of NELF-A (6-188) and NELF-C (183-590) was identified, crystallized and its structure solved to 2.8 Å resolution. Further, a novel sequence dependent interaction of NELF-AC with single-stranded nucleic acids was discovered and the architecture of the complete NELF complex elucidated by crosslinking coupled with mass-spectrometry. The results suggest a possible model for NELF action at the molecular level.
NELF, Negative Elongation Factor, promoter-proximal pausing, transcription, structural biology
Pöllmann, David
2015
English
Universitätsbibliothek der Ludwig-Maximilians-Universität München
Pöllmann, David (2015): Structural and functional studies on the negative elongation factor. Dissertation, LMU München: Faculty of Chemistry and Pharmacy
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Abstract

In many metazoans RNA polymerase II is stably paused after transcribing 20-60 nucleotides of a gene and awaits controlled release. The four subunit Negative Elongation Factor (NELF) is essential to this process which is called promoter-proximal pausing. In this work, a highly conserved NELF subcomplex consisting of NELF-A (6-188) and NELF-C (183-590) was identified, crystallized and its structure solved to 2.8 Å resolution. Further, a novel sequence dependent interaction of NELF-AC with single-stranded nucleic acids was discovered and the architecture of the complete NELF complex elucidated by crosslinking coupled with mass-spectrometry. The results suggest a possible model for NELF action at the molecular level.