Identification of a differentiation-inducible promoter in the E7 open reading frame of human papillomavirus type 16 (HPV-16) in raft cultures of a new cell line containing high copy numbers of episomal HPV-16 DNA

被引:129
作者
Grassmann, K
Rapp, B
Maschek, H
Petry, KU
Iftner, T
机构
[1] UNIV ERLANGEN NURNBERG, INST KLIN & MOLEK VIROL, D-91054 ERLANGEN, GERMANY
[2] HANNOVER MED SCH, INST PATHOL, HANNOVER, GERMANY
关键词
D O I
10.1128/JVI.70.4.2339-2349.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Gene expression of human papillomaviruses (HPV) is tightly linked to differentiation processes within the pluristratified epithelium. To analyze changes in the transcription pattern of HPV-16 during epithelial cell differentiation, we established a permanently growing HPV-16-positive cell Line, designated KG, from a vulvar intraepithelial neoplasm. KG cells of early passages harbored multiple copies of the HPV-16 DNA as episomes and were able to form a stratified epithelium in an organotypic raft culture system. Analysis of viral gene expression revealed the known transcription pattern of the early region of HPV-16 with the exception of a so far undefined mRNA class with start sites in the E7 open reading frame. Quantitative analysis of primer extension experiments with RNA from KG cells grown in monolayer and raft culture showed a strong induction of this transcript in differentiated KG cells, whereas the level of the mRNAs initiated at the early promoter P-97 remained almost constant. Primer extension analyses with four different primers and direct sequencing of the extension product revealed that the differentiation-inducible transcript initiated at a novel promoter with a major start site around nucleotide position 670 (P-670) in the E7 open reading frame of HPV-16 Sequence analysis of cDNAs derived from RNA of KG cells grown in raft culture suggested that the transcripts initiated at P-670 have a coding potential for an El boolean AND E4 fusion protein and for the E5 protein.
引用
收藏
页码:2339 / 2349
页数:11
相关论文
共 61 条
  • [1] STRUCTURAL AND TRANSCRIPTIONAL ANALYSIS OF HUMAN PAPILLOMAVIRUS TYPE-16 SEQUENCES IN CERVICAL-CARCINOMA CELL-LINES
    BAKER, CC
    PHELPS, WC
    LINDGREN, V
    BRAUN, MJ
    GONDA, MA
    HOWLEY, PM
    [J]. JOURNAL OF VIROLOGY, 1987, 61 (04) : 962 - 971
  • [2] DIFFERENTIAL PROMOTER UTILIZATION BY THE BOVINE PAPILLOMAVIRUS IN TRANSFORMED-CELLS AND PRODUCTIVELY INFECTED WART TISSUES
    BAKER, CC
    HOWLEY, PM
    [J]. EMBO JOURNAL, 1987, 6 (04) : 1027 - 1035
  • [3] SEQUENCE MICROHETEROGENEITY IN THE LONG CONTROL REGION OF CLINICAL ISOLATES OF HUMAN PAPILLOMAVIRUS TYPE-16
    BAVIN, PJ
    WALKER, PG
    EMERY, VC
    [J]. JOURNAL OF MEDICAL VIROLOGY, 1993, 39 (04) : 267 - 272
  • [4] AMPLIFICATION OF HUMAN PAPILLOMAVIRUS GENOMES INVITRO IS DEPENDENT ON EPITHELIAL DIFFERENTIATION
    BEDELL, MA
    HUDSON, JB
    GOLUB, TR
    TURYK, ME
    HOSKEN, M
    WILBANKS, GD
    LAIMINS, LA
    [J]. JOURNAL OF VIROLOGY, 1991, 65 (05) : 2254 - 2260
  • [5] PCR-BASED CDNA LIBRARY CONSTRUCTION - GENERAL CDNA LIBRARIES AT THE LEVEL OF A FEW CELLS
    BELYAVSKY, A
    VINOGRADOVA, T
    RAJEWSKY, K
    [J]. NUCLEIC ACIDS RESEARCH, 1989, 17 (08) : 2919 - 2932
  • [6] THE PREDOMINANT MESSENGER-RNA CLASS IN HPV16-INFECTED GENITAL NEOPLASIAS DOES NOT ENCODE THE E6 OR THE E7 PROTEIN
    BOHM, S
    WILCZYNSKI, SP
    PFISTER, H
    IFTNER, T
    [J]. INTERNATIONAL JOURNAL OF CANCER, 1993, 55 (05) : 791 - 798
  • [7] A NEW TYPE OF PAPILLOMAVIRUS DNA, ITS PRESENCE IN GENITAL CANCER BIOPSIES AND IN CELL-LINES DERIVED FROM CERVICAL-CANCER
    BOSHART, M
    GISSMANN, L
    IKENBERG, H
    KLEINHEINZ, A
    SCHEURLEN, W
    HAUSEN, HZ
    [J]. EMBO JOURNAL, 1984, 3 (05) : 1151 - 1157
  • [8] NORMAL KERATINIZATION IN A SPONTANEOUSLY IMMORTALIZED ANEUPLOID HUMAN KERATINOCYTE CELL-LINE
    BOUKAMP, P
    PETRUSSEVSKA, RT
    BREITKREUTZ, D
    HORNUNG, J
    MARKHAM, A
    FUSENIG, NE
    [J]. JOURNAL OF CELL BIOLOGY, 1988, 106 (03) : 761 - 771
  • [9] PHYLOGENETIC ANALYSIS OF 48 PAPILLOMAVIRUS TYPES AND 28 SUBTYPES AND VARIANTS - A SHOWCASE FOR THE MOLECULAR EVOLUTION OF DNA VIRUSES
    CHAN, SY
    BERNARD, HU
    ONG, CK
    CHAN, SP
    HOFMANN, B
    DELIUS, H
    [J]. JOURNAL OF VIROLOGY, 1992, 66 (10) : 5714 - 5725
  • [10] CHAN SY, 1992, J VIROL, V66, P571