Analysis of novel human papillomavirus type 16 late mRNAs in differentiated W12 cervical epithelial cells

The life cycle of human papillomavirus type 16 (HPV16) is intimately linked to differentiation of the epithelium it infects, and late events in the life cycle are restricted to the suprabasal layers. Here we have used 5'RACE of polyadenylated RNA isolated from differentiated W12 cells (cervical epithelial cells containing episomal copies of the HPV16 genome) that express virus late proteins to map virus late mRNAs. Thirteen different transcripts were identified. Extensive alternative splicing and use of two late polyadenylation sites were noted. A novel promoter located in the long control region was detected as well as P-97 and P-late. Promoters in the E4 and E5 open reading frames were active yielding transcripts where L1 or L2 respectively are the first open reading frames. Finally, mRNAs that could encode novel proteins E6 *(boolean AND)*E7,E6*(boolean AND)E4,E1(boolean AND)*E4 and E1(boolean AND)E2C (putative repressor E2) were identified, indicating that HPV16 may encode more late proteins than previously accepted. (c) 2006 Elsevier Inc. All rights reserved.