HelQ-BioID2 experiment to discover novel protein interactants of HelQ

ABSTRACT: HelQ is a 3’-5’ single stranded DNA (ssDNA) dependent helicase part of the Superfamily 2 helicases (SF2) which are characterised by the presence of two motor domains with a RecA-like fold which couples ATP hydrolysis to DNA translocation. The N-terminal domain of HelQ or N-HelQ (~300 amino acids) is non catalytically active, but sequence analysis has found a non-canonical PIWI-like motif (found in RNA splicing helicase Brr2) which is unable to bind DNA but is involved in displacement of RPA (Replication Protein A) from ssDNA 8. This study focussed on better understanding the role of this N-terminal domain of this helicase in vitro using BioID2. Biotin ligase protein interaction Identification generation 2 (BioID2) is a method for screening physiologically relevant protein interactions that occur in living cells originally developed by Roux et al.. BioID2 uses the promiscuous ability of Biotin ligase from A. aeolicus with a mutated catalytical domain (R40G) to biotinylate nearby proteins, these proteins can be pulled out of solution using streptavidin beads and protein identity discovered via LC-MS/MS. This method allows high throughput screening of the proteome surrounding HelQ and offers insight into potential DNA repair pathways involving HelQ.

INSTRUMENT(S): Q Exactive HF, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Bone Sarcoma

SUBMITTER: Jin Ke Michele Pan  

LAB HEAD: Panos Soultanas

PROVIDER: PXD049246 | Pride | 2025-05-06

REPOSITORIES: Pride