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BLM helicase protein negatively regulates stress granule formation through unwinding RNA G-quadruplex structures

ABSTRACT: We ran native 5% polyacrylamide-TBE gel for electro mobility shift assay (EMSA) for DNA/RNA G4 structures with or without recombinant human core-BLM (cBLM) protein, which was extracted and purified from E.coli in order to show binding of the RNA/DNA by this protein. To further evidence, we cut the equivalent gel area where the shifted band is located for all the wells (DNA only, DNA+cBLM, RNA, RNA+cBLM and cBLM only) and anlayzed by LC-MS/MS to identify human BLM and to see if there are some E.coli contaminations in those specific bands. Moreover, we analyzed the stock solution of the recombinant cBLM as general control.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Yehuda Danino

LAB HEAD: Eran Hornstein

PROVIDER: PXD042983 | Pride | 2023-10-24

REPOSITORIES: Pride

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BLM helicase protein negatively regulates stress granule formation through unwinding RNA G-quadruplex structures.

Danino Yehuda M YM Molitor Lena L Rosenbaum-Cohen Tamar T Kaiser Sebastian S Cohen Yahel Y Porat Ziv Z Marmor-Kollet Hagai H Katina Corine C Savidor Alon A Rotkopf Ron R Ben-Isaac Eyal E Golani Ofra O Levin Yishai Y Monchaud David D Hickson Ian D ID Hornstein Eran E

Nucleic acids research 20230901 17

Bloom's syndrome (BLM) protein is a known nuclear helicase that is able to unwind DNA secondary structures such as G-quadruplexes (G4s). However, its role in the regulation of cytoplasmic processes that involve RNA G-quadruplexes (rG4s) has not been previously studied. Here, we demonstrate that BLM is recruited to stress granules (SGs), which are cytoplasmic biomolecular condensates composed of RNAs and RNA-binding proteins. BLM is enriched in SGs upon different stress conditions and in an rG4-d ...[more]

PMID: 37503837

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