Products/Services Used | Details | Operation |
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Recombinant Antibody Services> | , abstracts N/A Dynabeads Human T-Activator CD3/CD28 Life Technologies Cat#11131D Histone H4, Human, Recombinant New England Biolabs Cat#M2504S Histone H4 Peptide aa 1–21 Cayman Chemical Cat#10854 Histone H4 Peptide aa 15–24 Cayman Chemical Cat#10380 Histone H4 Peptide aa 1–21 K20A GenScript Cat#U1417CA160_3 SMYD2 Human Recombinant Protein Sigma-Aldrich Cat#SRP0151 SMYD2 Y240F Human Recombinant Protein Active Motif Cat#31324 Critical Commercial Assays EasySepHuman CD4+ T Cell Isolation Kit STEMCELL Technologies Cat#19052 RosetteSep Human CD4+ T Cell Enrichment Cocktail STEMCELL Technologies Cat#15062 RNeasy Plus Mini Kit QIAGEN Cat#74136 Luciferase Assay System Promega Cat#E4030 CellTiter-Blue Cell Viability Assay Promega Cat#G8081 ApoTox-Glo Triplex Assays Promega Cat#G6321 Experimental Models: Cell Lines HEK293T ATCC CRL-3216; RRID: CVCL_0063 J-Lat A2, male, derived from Jurkat cells Jordan et al.... For reactions, 2 mg of histones (isolated from HEK293T cells), recombinant histone 4 (New England Biolabs), synthetic histone 4 aa 1–21 and aa 15–24 peptides (Cayman Chem- ical), or synthetic histone H4 aa 1–21 with a K20A mutation (GenScript) were incubated with recombinant WT SMYD2 (Sigma-Aldrich) or SMYD2 Y240F (Active Motif) in a buffer containing 50 mM Tris-HCl, pH 9, 0. | Get A Quote |
Transcriptional latency of HIV is a last barrier to viral eradication. Chromatin-remodeling complexes and post-translational histone modifications likely play key roles in HIV-1 reactivation, but the underlying mechanisms are incompletely understood. We performed an RNAi-based screen of human lysine methyltransferases and identified the SET and MYND domain-containing protein 2 (SMYD2) as an enzyme that regulates HIV-1 latency. Knockdown of SMYD2 or its pharmacological inhibition reactivated latent HIV-1 in T cell lines and in primary CD4 T cells. SMYD2 associated with latent HIV-1 promoter chromatin, which was enriched in monomethylated lysine 20 at histone H4 (H4K20me1), a mark lost in cells lacking SM... More