title: HMGB1 gene knockout in mouse embryonic fibroblasts results in reduced telomerase activity and telomere dysfunction.
abstract: Telomere repeats are added onto chromosome ends by telomerase, consisting of two main core components: a catalytic protein subunit (telomerase reverse trancriptase, TERT), and an RNA subunit (telomerase RNA, TR). Here, we report for the first time evidence that HMGB1 (a chromatin-associated protein in mammals, acting as a DNA chaperone in transcription, replication, recombination, and repair) can modulate cellular activity of mammalian telomerase. Knockout of the HMGB1 gene (HMGB1 KO) in mouse embryonic fibroblasts (MEFs) results in chromosomal abnormalities, enhanced colocalization of γ-H2AX foci at telomeres, and a moderate shortening of telomere lengths. HMGB1 KO MEFs also exhibit significantly (>5-fold) lower telomerase activity than the wild-type MEFs. Correspondingly, enhanced telomerase activity is observed upon overexpression of HMGB1 in MEFs. HMGB1 physically interacts with both TERT and TR, as well as with active telomerase complex in vitro. However, direct interaction of HMGB1 with telomerase is most likely not accountable for the observed higher telomerase activity in HMGB1-containing cells, as revealed from the inability of purified HMGB1 protein to stimulate telomerase activity in vitro. While no transcriptional silencing of TERT is observed in HMGB1 KO MEFs, levels of TR are diminished (~3-fold), providing possible explanation for the observed lower telomerase activity in HMGB1 KO cells. Interestingly, knockout of the HMGB2 gene elevates telomerase activity (~3-fold) in MEFs, suggesting that the two closely related proteins of the HMGB family, HMGB1 and HMGB2, have opposite effects on telomerase activity in the cell. The ability of HMGB1 to modulate cellular activity of telomerase and to maintain telomere integrity can help to understand some aspects of the protein involvement in chromosome stability and cancer.
authors.affiliation: Academy of Sciences of the Czech Republic, Institute of Biophysics, Brno, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Polanská
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Dobšáková
authors.forename: Martina
authors.initials: M
authors.lastname: Dvořáčková
authors.forename: Jiří
authors.initials: J
authors.lastname: Fajkus
authors.forename: Michal
authors.initials: M
authors.lastname: Štros
mesh_terms: D000818:Animals; D002460:Cell Line; D002869:Chromosome Aberrations; D004249:DNA Damage; D053938:DNA Fragmentation; D004261:DNA Replication; D015536:Down-Regulation; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D055786:Gene Knockout Techniques; D024243:HMGB1 Protein / Q000235:genetics* / Q000378:metabolism; D024261:HMGB2 Protein / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008856:Microscopy, Fluorescence; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D016615:Telomere / Q000378:metabolism / Q000473:pathology
chemical_list: D024243:HMGB1 Protein; D024261:HMGB2 Protein; D006657:Histones; C475961:gamma-H2AX protein, mouse; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase; C509187:Tert protein, mouse