taip induces apoptosis in hepatocellular carcinoma-derived cells klaas kooistra biological...
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![Page 1: TAIP induces apoptosis in hepatocellular carcinoma-derived cells Klaas Kooistra Biological Chemistry, Molecular Genetics LIC, Leiden University LC Workshop](https://reader030.vdocuments.net/reader030/viewer/2022033104/56649d225503460f949f8cee/html5/thumbnails/1.jpg)
TAIP induces apoptosis in hepatocellular carcinoma-derived cells
Klaas Kooistra
Biological Chemistry, Molecular GeneticsLIC, Leiden University
LC Workshop Proteins killing Tumour CellsNovember 1, 2007
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A specific TT virus strain has a CAV-like genome
TTV protein (TAIP) harbors apoptin-like characteristics?• Amino-acid sequence similarities?• Apoptosis?
TAIP-studies in human tumor cell lines
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![Page 3: TAIP induces apoptosis in hepatocellular carcinoma-derived cells Klaas Kooistra Biological Chemistry, Molecular Genetics LIC, Leiden University LC Workshop](https://reader030.vdocuments.net/reader030/viewer/2022033104/56649d225503460f949f8cee/html5/thumbnails/3.jpg)
Specific TT virus (TTV) strain
TT viruses belong to human circoviridae• Single stranded circular DNA virus
Within TT viruses a very high genetic variability exists
TTV strain (GenBank: AB008394) described by Miyata• Isolated from Japanese patient with non-A-E hepatitis
Attracted our specific attention for …..
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Miyata et al. 1999
Apoptin
TAIP
GC-rich region
SP-1 site
Poly(A) site
TATA box
TTV-strain has CAV-like genomic organization
TTV: torque teno virusCAV: chicken anemia virus
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TAIP
Apt
TAIP
Apt
TAIP
Apt
TAIP amino acid sequence shows “similarities” with apoptin’s
Hydrophobic a.a. stretches; Proline distribution; Thr/Ser-rich
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Expression of TAIP in human tumor cells
Putative TTV-derived TAIP of 105 a.a.
Approach0.36 kb Myc-TAIP DNA was synthesized (Midland, USA) Cloned in pCMV-expression vector
15 kD
TAIP production in COS-1 cells
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TAIP
IFA / HUH-7 cells
TAIP DNA
apoptin apoptin DNA
Cytoplasmic TAIP inducesapoptosis in HUH-7 cells
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Transient transfection of three human hepatocellular carcinoma (HCC) cell lines
TAIP
Apoptin
Lac Z
5 days post transfection
TAIP induces apoptosis human HCC cells
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Transient transfection of three non-HCC cell lines
TAIP induces much less apoptosis in non-HCC
TAIP
Apoptin
Lac Z
5 days post transfection
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TAIP-induces apoptosis in HCC significantlymore than in non-HCC
TAIP
Apoptin
Lac Z
Preliminary results: TAIP does not induce apoptosisin normal rat hepatocytes
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TAIP induces p53 independent apoptosis
TAIP induces apoptosis preferentially in HCC cells
TAIP is localized in the cytoplasm in cancer cells
TAIP non-toxic for normal cells (?)
Conclusions
TAIP functions differently than apoptin?!
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Acknowledgements
Ying-Hui ZhangNiek HendriquezBertram Weiss
Dominik MumbergMathieu Noteborn
Thank you for your attention
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1. Miyata et al., Identification of a novel GC-rich 113-nucleotide region to complete the circular, single-stranded DNA genome of TT virus, the first human circovirus.J Virol. 1999 May;73(5):3582-6.
2. Kooistra et al., TT virus-derived apoptosis-inducing protein induces apoptosis preferentially in hepatocellular carcinoma-derived cells.J Gen Virol. 2004 Jun;85(Pt 6):1445-50.
3. Hino and Miyata, Torque teno virus (TTV): current status.Rev Med Virol. 2007 Jan-Feb;17(1):45-57.
Literature of interest