BLM解旋酶N端301~650肽段互作蛋白的筛选及鉴定

doi:10.3969/j.issn.1674-7968.2026.04.013

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Abstract As a vital member of the human recombination Q (RecQ) helicase family, Bloom syndrome mutated protein (BLM) plays essential roles in multiple cellular metabolic processes, including DNA replication, repair, recombination, transcription, and telomere maintenance. Structural studies on BLM helicase have revealed that, in addition to the 3 canonical conserved domains characteristic of the RecQ helicase family—the helicase domain (HD), RecQ C-terminal domain (RecQ-Ct) and helicase and RNase D C-terminal domain (HRDC)—a nuclear localization signal (NLS) is also identified at its carboxyl terminus (C-terminus).However, research on its amino terminus (N-terminus) remains limited. This research group's structural analysis of the BLM N-terminus has found that a large number of threonine phosphorylation sites were present in the region spanning 619~843 bp. It was hypothesized that these sites play a critical function in the post-translational modifications (PTMs) of BLM helicase, yet the cellular proteins involved in this physiological process remain incompletely elucidated. To screen for cellular proteins that interact with the N-terminal 301~650 peptide fragment of BLM helicase (BLM_301~650) and further characterize their interaction with the BLM N-terminus, this present study employed immunoprecipitation-high-performance liquid chromatography tandem mass spectrometry (IP-HPLC-MS/MS) to identify BLM_301~650-interacting proteins in the prostate cancer (PC3) cell line. Bioinformatic analyses of the screened interacting proteins were performed using the database for annotation, visualization and integrated discovery (DAVID) and search tool for the retrieval of interacting genes/proteins (STRING). The subcellular colocalization between BLM_301~650 and its interacting proteins was validated via immunofluorescence assay, while the interaction relationship was confirmed by Pull-down assays. IP-HPLC-MS/MS results demonstrated that a total of 172 proteins specifically binding to BLM_301~650 were identified. KEGG pathway enrichment analysis indicated that these interacting proteins were predominantly enriched in the ribosome pathway. Protein-protein interaction (PPI) network analysis further identified ribosomal protein S27a (RPS27A) as a key interacting factor of BLM_301~650. Immunofluorescence and Pull-down assays verified that BLM_301~650 and RPS27A colocalized in the nucleoplasm and exhibited a direct targeting relationship. Collectively, the direct interaction between RPS27A and the N?terminal region of BLM helicase serves as a novel molecular bridge for the crosstalk between the ribosomal pathway and the DNA repair pathway, and is of great scientific significance for elucidating the mechanisms underlying genome stability maintenance, tumorigenesis, and DNA damage response.

Key words： BLM helicase N-terminal domain Proteomics Ribosomal protein S27a (RPS27A) Ribosomal pathway

Received: 03 November 2025

ZTFLH:

S857.4

Corresponding Authors: ^*jfzhao@gzu.edu.cn

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	Articles by authors
	ZHAO Jia-Fu
	YANG Yuan-Qing
	WEN Xiao-Yan
	ZHANG Xi-Lu
	CHEN Yi
	PAN Zhi-Ren
	WEI Zhong-Yi

Cite this article:

ZHAO Jia-Fu,YANG Yuan-Qing,WEN Xiao-Yan, et al. Screening and Identification of Proteins Interacting with the N-terminal 301~650 Peptide Fragment of BLM Helicase[J]. 农业生物技术学报, 2026, 34(4): 834-843.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2026.04.013 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2026/V34/I4/834

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