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. 2005 Jul;79(14):9301-5.
doi: 10.1128/JVI.79.14.9301-9305.2005.

Cloning and identification of a microRNA cluster within the latency-associated region of Kaposi's sarcoma-associated herpesvirus

Mark A Samols  1 , Jianhong Hu, Rebecca L Skalsky, Rolf Renne
Affiliations

Cloning and identification of a microRNA cluster within the latency-associated region of Kaposi's sarcoma-associated herpesvirus

Mark A Samols et al. J Virol. 2005 Jul.

Abstract

MicroRNAs (miRNAs) are small, noncoding regulatory RNA molecules that bind to 3' untranslated regions (UTRs) of mRNAs to either prevent their translation or induce their degradation. Previously identified in a variety of organisms ranging from plants to mammals, miRNAs are also now known to be produced by viruses. The human gammaherpesvirus Epstein-Barr virus has been shown to encode miRNAs, which potentially regulate both viral and cellular genes. To determine whether Kaposi's sarcoma-associated herpesvirus (KSHV) encodes miRNAs, we cloned small RNAs from KSHV-positive primary effusion lymphoma-derived cells and endothelial cells. Sequence analysis revealed 11 isolated RNAs of 19 to 23 bases in length that perfectly align with KSHV. Surprisingly, all candidate miRNAs mapped to a single genomic locale within the latency-associated region of KSHV. These data suggest that viral and host cellular gene expression may be regulated by miRNAs during both latent and lytic KSHV replication.

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Figures

FIG. 1.
FIG. 1.
Identification of an miRNA cluster in the intragenic region between K12 and ORF71. (A) Schematic of the latency-associated region of KSHV. All nucleotide annotations are based on the BC-1 KSHV sequence (24). ORFs, transcripts, DR1 and DR2, and promoters are drawn as described in references , , , and . The miRNA cluster, as well as the proposed kaposin transcripts by Li et al. (17), are shown in orange. (B) Sequences of cloned candidate miRNA species, nucleotide position, and cloning frequency are indicated. Preliminary names of candidate miRNAs as submitted to GenBank are given (accession number AY973824).
FIG. 2.
FIG. 2.
Isolated miRNAs fold into hairpin structures of appropriate length. (A) Secondary structure predictions of candidate KSHV miRNA sequences using mFold (28). Shown are only structures with the lowest free energy. (B) Northern blot analysis for the expression of KSHV miRNAs 3-3p, 4, and 10 and the human miRNA hsa-miR-16 as a positive control. Thirty micrograms of total RNA from BCBL-1, TPA-induced BCBL-1, TIVE-LTC, and latently infected SLK cells (SLK KS+) was separated on 15% PAGE, blotted to nylon membranes, probed with an end-labeled antisense oligonucleotide, and detected by phosphorimaging as previously described (16).
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