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Structural requirements for photo-induced RNA-protein cross-linking

ABSTRACT: Cross-linking of isotope-labelled RNA coupled with mass spectrometry (CLIR-MS) was used to study the UV cross-linking behavior of in vitro reconstituted FOX1 RRM in complex with its cognate RNA sequence, the Fox binding element (FBE), (U)GCAUGU. The FBE heptanucleotide was subsequently mutated, and the impact on UV cross-linking and affinity investigated. Cross-linking was performed using irradiation under 254 nm light, relying on the inherent reactivity of ribonucleotides.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Christopher Sarnowski

LAB HEAD: Alexander Leitner

PROVIDER: PXD031381 | Pride | 2022-06-09

REPOSITORIES: Pride

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			Action	DRS
	200625_LU2_csarnowski_AK301_01.raw	Raw
	200625_LU2_csarnowski_AK301_06.raw	Raw
	Fig1b.csv	Csv
	Fig1c.csv	Csv
	Fig2c_f.csv	Csv

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Publications

Nucleotide-amino acid π-stacking interactions initiate photo cross-linking in RNA-protein complexes.

Knörlein Anna A Sarnowski Chris P CP de Vries Tebbe T Stoltz Moritz M Götze Michael M Aebersold Ruedi R Allain Frédéric H-T FH Leitner Alexander A Hall Jonathan J

Nature communications 20220517 1

Photo-induced cross-linking is a mainstay technique to characterize RNA-protein interactions. However, UV-induced cross-linking between RNA and proteins at "zero-distance" is poorly understood. Here, we investigate cross-linking of the RBFOX alternative splicing factor with its hepta-ribonucleotide binding element as a model system. We examine the influence of nucleobase, nucleotide position and amino acid composition using CLIR-MS technology (crosslinking-of-isotope-labelled-RNA-and-tandem-mass ...[more]

PMID: 35581222

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