We have concentrated
on identifying tumor suppressor genes using a functional
genetic approach in mammalian cell culture models. This
required the development and employment of a powerful
new genetic method that can be used to generate and
identify recessive alterations. This method is termed
technical knockout (TKO). It is based on random inactivation
of genes mediated by expression of an anti-sense cDNA
library, followed by a selection for a specific phenotypic
alteration and identification of the specific anti-sense
construct responsible for the phenotypic change. We
have successfully used TKO selection to identify several
genes involved in two discrete phenotypic alterations
that are often associated with tumors–loss of
sensitivity to programmed cell death (PCD) and loss
of sensitivity to contact mediated growth arrest.
To date, fourteen genes involved in PCD have been cloned
by using TKO. While analysis of these genes has just
begun, it was found that one gene, Dap-kinase, is frequently
lost in human tumors, as expected for tumor suppressor
genes. Another gene, Cathepsin D, has been linked to
a pathway involved in autoimmunity. We are now extensively
studying the cloned genes, and we expect to demonstrate
their critical roles in various PCD functions, such
as organ formation; immune and nervous system homeostasis;
viral defense; and heart, liver and kidney failure,
as well as in tumor suppression.
Selected Publications:
Frenkel, N., Spaete, R.R., Vlazny, D.A., Deiss, L.P.,
and Locker, H. The herpes simplex virus amplicon—A
novel animal-virus cloning vector. In: Eukaryotic Viral
Vectors, Y. Gluzman, ed. Cold Spring Harbor Laboratory,
New York, pp. 205-209, 1982.
Frenkel, N., Deiss, L.P., and Spaete, R.R. Studies of
cis acting replication functions of herpes simplex virus
DNA. J. Cell. Biochem. Supplement 8B, p. 198, 1984.
Frenkel, N., Deiss, L.P., Kwong, A.D., and Spaete, R.R.
Defective virus vectors (amplicons) derived from herpes
simplex viruses. In: Gene Transfer and Cancer, M.L.
Pearson and N.L. Sternberg, eds. Raven Press, New York,
pp. 205-113, 1984.
Frenkel, N., Deiss, L.P., and Spaete, R.R. Studies of
HSV DNA propagation using HSV amplicons. In: Herpesvirus,
F. Rapp, ed. Alan R. Liss, Inc., New York,
pp. 289-299, 1984.
Mocarski, E.S., Deiss, L.P., and Frenkel, N. Nucleotide
sequence and structural features of a novel US-a junction
present in a defective herpes simplex virus genome.
J. Virol. 55: 140-146, 1985.
Deiss, L.P., and Frenkel, N. Herpes simplex virus amplicon:
Cleavage of concatemeric DNA is linked to packaging
and involves amplification of the terminally reiterated
a sequence. J. Virol. 57: 933-941, 1986.
Manservigi, R., Cassai, E., Deiss, L.P., Di Luca, D.,
Segala, V., and Frenkel, N. Sequences homologous to
two separate transforming regions of herpes simplex
virus DNA are linked in two human genital tumors. Virology
155: 192-201, 1986.
Deiss, L.P., Chou, J., and Frenkel, N. Functional domains
within the a sequence involved in the cleavage/packaging
of herpes simplex virus DNA. J. Virol. 59: 605-618,
1986.
Deiss, L.P., and Kimchi, A. A genetic tool used to identify
Thioredoxin as a mediator of a growth inhibitory signal.
Science 252: 117-120, 1991.
Deiss, L.P., Feinstein, E., Berissi, H., Cohen, O.,
and Kimchi, A. Identification of a novel serine/threonine
kinase and a novel 15-kD protein as potential mediators
of the g interferon-induced cell death. Genes Dev. 9:
15-30, 1995.
Kissil, J.L., Deiss, L.P., Bayewitch, M., Raveh, T.,
Khaspekov, G., and Kimchi, A. Isolation of DAP3: a novel
mediator of interferon-g-induced cell death. J. Biol.
Chem. 270: 27932-27936, 1995.
Deiss, L.P., Galinka, H., Berissi, H., Cohen, O., and
Kimchi, A. Cathepsin D protease mediates programmed
cell death induced by interferon-g, Fas/APO-1 and TNF-a.
EMBO J. 15(15): 3861-3870, 1996.
Levy-Strumpf, N., Deiss, L.P., Berissi, H., and Kimchi,
A. DAP-5, a novel homolog of eukaryotic translation
initiation factor 4G isolated as a putative modulator
of gamma interferon-induced programmed cell death. Mol.
Cell. Biol. 17(3): 1615-1625, 1997.
Gudkov, A.V., Roninson, I.G., Brown, R., Kimchi, A.,
Cohen, O., Kissil, J., Raveh, T., Inbal, B., Levy-Strumpf,
N., Berissi, H., and Deiss, L.P. Functional approaches
to gene isolation in mammalian cells. Science, 285:
299, 1999.
Yehiely, F., and Deiss L.P. Apoptosis and cancer. In:
Basic science of cancer, Kruh, G.D., and Tew, K.D.,
eds. Current Medicine, Inc., Philadelphia, pp. 216-232,
2000.
Kotlo, K.U., Yehiely, F., Efimova, E., Harasty, H.,
Hesabi, B., Shchors, K., Einat, P., Rozen, A., Berent,
A., and Deiss, L.P. Nrf2 is an inhibitor of the Fas
pathway as identified by Achilles Heel Method, a new
function-based approach to gene identification in human
cells. (Submitted)
Kotlo, K.U., Yehiely, F., Sonenberg, N., and Deiss,
L.P. Dap-5 regulates the level of a tumor susceptibility
gene by directly binding to its 5' UTR. (Manuscript
in preparation)
Cilensek, Z.M., Yehiely, F., Kular, R.K., and Deiss,
L.P. A member of the GAGE family of tumor antigens is
an anti-apoptotic gene that confers resistance to Fas/CD95/APO-1,
Interferon-g, Taxol and g-irradiation. Cancer Biology
and Therapy. (In press)
Shchors, K., Yehiely, F., and Deiss, L.P. Expression
of the 3’UTR of DAP-7 protects cells from Interferon-g
induced apoptosis and regulates the stability of p21/Cip-1/SDI-1/WAF-1.
(Manuscript in preparation)
Shchors, K., Yehiely, F., Brewer, G., and Deiss, L.P.
Cell death inhibiting RNA (CDIR) derived from a 3' untranslated
region binds AUF1 and heat shock protein 27. J. Biol.
Chem. (Manuscript under revision)
Bader, H., Yehiely, F., Gottlieb, K., Hay, N., and Deiss,
L.P. An inhibitor identified by Achilles Heel Method
preferentially kills transformed cells. (Manuscript
in preparation) |
