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oai:figshare.com:article/34049126 2026-10-01T17:42:56Z category_4 category_12 category_13 category_14 category_64 category_128 category_132 category_134 portal_5 item_type_1 month_year_10_2026

MARCHF5 requires its ubiquitin ligase and dimerization activities in PEL. Prasanth Viswanathan (10314782) Justine R. Bersonda (25157885) Jackson Gill (25157888) Alyssandra Navarro (25157891) Allison C. Farrar (25157894) Thomas Caldarera (25157897) Andrew Murphy (341397) Daniel Dunham (25157900) Karl W. Boehme (11384016) Mark Manzano (25157903) Biochemistry Cell Biology Genetics Molecular Biology Cancer Hematology Infectious Diseases Virology systems biology approach regulating mitochondrial homeostasis primary effusion lymphoma immunoprecipitation experiments show endogenous reciprocal co emerging clinical relevance cell viability due newly appreciated interaction causes several malignancies unexpected genetic interaction findings thus provide div >< p marchf5 oncogenes may pel cells suggesting mcl1 motivated us genetic interaction tumor cells causes resistance thus investigated therapeutic potential survival role significant increase protein complex previously shown pel ). mechanistic link kaposi ’ fitness advantage e3 ligase drug development dimerization activities crispr knockout combination therapies chemotherapy regimens attractive target associated herpesvirus apoptotic signal apoptotic activities antiviral signaling A. Western blots of endogenous and 3XFLAG-tagged MARCHF5. BCBL-1 iCas9 cells expressing gRNAs targeting MARCHF5 or AAVS1 were transduced with lentiviruses constitutively expressing cDNAs for mCherry, codon-optimized MARCHF5 (coMARCHF5) wildtype, H43W ligase mutant, or 4GL dimerization mutant and treated with Doxycycline (Dox) (0.5 μg/mL) for 9 days. To visualize the coMARCHF5 expressed from the lentivirus, cells were treated with 2.5 μM MG132 for 8 hour prior to harvesting for Western blotting. B. Quantification of Western blots in panel A normalized to total protein stain and gAAVS1. C. Cumulative live cell counts (relative to control gAAVS1) of BCBL-1 iCas9 cells in panel A after the addition of Dox (no MG132). Cells were seeded at 3 x 105 cells/mL and Cas9 expression was induced by Dox treatment (0.5 μg/mL). Every three days, total live cells were counted by trypan blue exclusion assay and split back to 3 x 105 cells/mL to allow the cells to continue proliferating. Cumulative live cell counts are expressed relative to the negative control gRNA AAVS1. Statistical differences were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (n = 6-8). Adjusted p values reflect post-hoc comparisons between gAAVS1 to either MARCHF5 gRNA at the specific time point. Error bars, standard error of the mean. 2026-10-01T17:54:28Z Image Figure 10.1371/journal.ppat.1014665.g004 https://figshare.com/articles/figure/_p_MARCHF5_requires_its_ubiquitin_ligase_and_dimerization_activities_in_PEL_p_/34049126 CC BY 4.0