Identification of conserved hydrophobic C-terminal residues of the human papillomavirus type 1 E1∧E4 protein necessary for E4 oligomerisation in vivo

被引:11
作者
Ashmole, I [1 ]
Gallimore, PH [1 ]
Roberts, S [1 ]
机构
[1] Univ Birmingham, Sch Med, Canc Res Campaign, Inst Canc Studies, Birmingham B15 2TA, W Midlands, England
关键词
D O I
10.1006/viro.1997.8909
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Previous studies have shown that human papillomavirus (HPV) E4 proteins undergo oligomerisation, although the precise sequences involved have not been identified. Using the yeast two-hybrid system we have identified HPV 1 E4 sequences that are critical to multimerisation. Fusion proteins were created by linking wild-type and mutant E4 proteins to a LexA DNA-binding domain or a B42 transactivation domain. HPV 1 E4:E4 interactions were examined by expression of these fusion proteins in Saccharomyces cerevisiae. This assay showed that (1) amino acid residues 95 to 115 at the carboxy-terminus were critical for oligomerisation and (2) hydrophobic residues (isoleucine 107, phenylalanine 114) in this domain are major determinants in the formation of oligomers. Interestingly, the carboxy-terminal domain shares homology with other E4 proteins of cutaneous HPV types and, furthermore, positions 107 and 114 are conserved residues. Substitution of the conserved aspartate amino acids (residues 110 and 112) did not abrogate 54 oligomerisation. Chemical cross-linking of wart and recombinant (baculovirus-expressed) HPV 1 E4 protein indicated that in solution this viral protein forms complexes consistent in size with either trimers or tetramers. These complexes were resistant to urea denaturation and are not dependent on the formation of disulphide linkages. A mutant protein containing a deletion of residues 110 to 115 was unable to form oligomers following cross-linking supporting a role for this region in mediating E4:E4 interactions. We conclude that oligomerisation of the HPV 1 E4 protein is likely to be mediated by carboxy-terminal residues and that conserved hydrophobic residues of this domain play a major role in E4 oligomerisation. (C) 1998 Academic Press.
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页码:221 / 231
页数:11
相关论文
共 35 条
  • [1] 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
  • [2] BREITBURD F, 1987, PAPILLOMAVIRUSES CAN, P115
  • [3] HUMAN PAPILLOMAVIRUS TYPE-11 E1-BOOLEAN-AND-E4 AND L1 PROTEINS COLOCALIZE IN THE MOUSE XENOGRAFT SYSTEM AT MULTIPLE TIME POINTS
    BROWN, DR
    BRYAN, JT
    PRATT, L
    HANDY, V
    FIFE, KH
    STOLER, MH
    [J]. VIROLOGY, 1995, 214 (01) : 259 - 263
  • [4] IDENTIFICATION AND MAPPING OF HUMAN PAPILLOMAVIRUS TYPE-1 RNA TRANSCRIPTS RECOVERED FROM PLANTAR WARTS AND INFECTED EPITHELIAL-CELL CULTURES
    CHOW, LT
    REILLY, SS
    BROKER, TR
    TAICHMAN, LB
    [J]. JOURNAL OF VIROLOGY, 1987, 61 (06) : 1913 - 1918
  • [5] HUMAN PAPILLOMAVIRUS TYPE-6 AND TYPE-11 MESSENGER-RNAS FROM GENITAL CONDYLOMATA ACUMINATA
    CHOW, LT
    NASSERI, M
    WOLINSKY, SM
    BROKER, TR
    [J]. JOURNAL OF VIROLOGY, 1987, 61 (08) : 2581 - 2588
  • [6] Croissant O, 1985, Clin Dermatol, V3, P43, DOI 10.1016/0738-081X(85)90048-3
  • [7] COEXPRESSION OF THE HUMAN PAPILLOMAVIRUS TYPE-16 E4 AND L1 OPEN READING FRAMES IN EARLY CERVICAL NEOPLASIA
    CRUM, CP
    BARBER, S
    SYMBULA, M
    SNYDER, K
    SALEH, AM
    ROCHE, JK
    [J]. VIROLOGY, 1990, 178 (01) : 238 - 246
  • [8] ANALYSIS OF HPV-1 E4 GENE-EXPRESSION USING EPITOPE-DEFINED ANTIBODIES
    DOORBAR, J
    EVANS, HS
    CONERON, I
    CRAWFORD, LV
    GALLIMORE, PH
    [J]. EMBO JOURNAL, 1988, 7 (03) : 825 - 833
  • [9] SEQUENCE DIVERGENCE YET CONSERVED PHYSICAL CHARACTERISTICS AMONG THE E4 PROTEINS OF CUTANEOUS HUMAN PAPILLOMAVIRUSES
    DOORBAR, J
    CONERON, I
    GALLIMORE, PH
    [J]. VIROLOGY, 1989, 172 (01) : 51 - 62
  • [10] IDENTIFICATION OF THE HUMAN PAPILLOMA VIRUS-1A E4 GENE-PRODUCTS
    DOORBAR, J
    CAMPBELL, D
    GRAND, RJA
    GALLIMORE, PH
    [J]. EMBO JOURNAL, 1986, 5 (02) : 355 - 362