More studies from recent reports indicate miR-K12-5, miR-K12-9 and miR-K12-10 repression of Bcl-2-associated transcription factor 1, an apoptosis-inducing factor (238), miR-K12-3 reduction of LRRC8D (leucine rich repeat containing 8 family, member D), a leucin-rich type III transmembrane protein involved in proliferation and activation of lymphocytes, NHP2L1 (non-histone chromosome protein 2-like 1) (239), and a transcription factor NFIB (240), miR-K12-1 inhibition of p21 241 and NFkappaB inhibitor IkappaBalpha (242), miR-K12-4-3p suppression of GEMIN8 (239), miR-K12-10a reduction of TWEAKR (243), miR-K12-1, -6, and -11 decrease of MAF (musculoaponeurotic fibrosarcoma oncogene homolog) (244), K12-4-5p targeting retinoblastoma (Rb)-like protein 2 (Rbl2) and miR-K12-5 and -9*suppressing ORF50 mRNA (245,246).
We identified a high confidence interaction site for kshv-miR-K12-4-3p in the coding sequence of GEMIN8 and for kshv-miR-K12-3 in the coding sequence of NHP2L1.
Currently, the validated targets of KSHV miRNAs (Table 1) represent a collection of genes identified by unbiased methods or chosen by previously identified functions.Table 1 KSHVmiRNA validated targets. All targets have been validated using ectopic expression of miRNAs to demonstrate that a luciferase reporter or ectopic reporter are repressed. Additional validations include(1) exogenous reporters have been mutated to disrupt targeting by a ectopic miRNA,(2) ectopic miRNA suppresses endogenous target mRNA and/or protein levels,(3) de novo infection with KSHV represses endogenous target expression,(4) miRNA inhibitors and/or mutant virus display derepression of endogenous target mRNA and/or protein levels, and(5) repression of miRNA target is observed in KSHV clinical samples.
