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J. Biol. Chem., Vol. 277, Issue 17, 14547-14556, April 26, 2002

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Kaposi's Sarcoma-associated Herpesvirus K8 Exon 3 Contains Three 5'-Splice Sites and Harbors a K8.1 Transcription Start Site^*

Shuang Tang and Zhi-Ming Zheng

From the HIV and AIDS Malignancy Branch, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892

Kaposi's sarcoma-associated herpesvirus (KSHV) K8 and K8.1 open reading frames are juxtaposed and span from nucleotide (nt) 74850 to 76695 of the virus genome. A K8 pre-mRNA overlaps the entire K8.1 coding region, and alternative splicing of KSHV K8 and K8.1 pre-mRNAs each produces three isoforms (, , and ) of the mRNAs. We have mapped the 5' end of the K8.1 RNA in butyrate-induced KSHV-positive JSC-1 cells to nt 75901 in the KSHV genome and have shown that exon 3 of the K8 pre-mRNA in JSC-1 cells covers most part of the intron 3 defined previously and has three 5'-splice sites (ss), respectively, at nt 75838, 76155, and 76338. Selection of the nt 75838 5'-ss dictates the K8 mRNA production and overwhelms the RNA processing. Alternative selection of other two 5'-ss is feasible and leads to production of two additional bicistronic mRNAs, K8/K8.1 and -. However, the novel bicistronic K8/K8.1 mRNAs translated a little K8 and no detectable K8.1 proteins in 293 cells. Data suggest that production of the K8/K8.1 mRNAs may be an essential way to control K8 mRNAs, especially K8, to a threshold at RNA processing level.

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