MOLECULAR CHARACTERIZATION OF PERFORIN GENE REGULATION

Project: Research project

Project Details

Description

DESCRIPTION (Adapted from Investigator's abstract): Natural killer (NK)
cells and cytotoxic T lymphocytes (CTL) defeat immunogenic tumors and virus
infections. Physiologically, most CTL reside in the body as pre-effectors
(pre-CTL) that are unable to kill. When the immune system is confronted
with intracellular pathogens, preCTL are expanded and armed. During this
process many genes are newly expressed or upregulated in preCTL, leading to
their proliferation and differentiation into effector CTL(eff CTL). One of
these genes is perforin, a membrane "perforating" killer molecule essential
for the function of the adaptive immune system. In contrast, NK cells,
perhaps best thought of as primordial CTL that lack receptors for a specific
antigen, constitutively express perforin and kill without activation. Our
experiments have shown that perforin gene expression is determined at the
transcriptional level and that this process requires regulatory domains in
addition to the promoter. This application will continue to identify and
characterize these regulatory domains by elucidating constitutive and
inducible nuclear events at the perforin gene locus in NK cells and CTL.
Molecules involved in this process ultimately may be of therapeutical use or
may become therapeutical targets. From the immunological and developmental
point of view, the regulatory domains ultimately may shed light on
transcription factors that control not only the perforin gene but also the
commitment to and progression of the perforin expressing lineages during
hematolymphopoiesis. This application specifically proposes to: 1) explore
the presence of additional regulatory domains within the boundaries of an
active perforin gene locus in tumor cell lines; 2) characterize the function
of potential regulatory domains in tumor cell lines; and 3) demonstrate the
function of the regulatory domains in normal cells. To achieve these aims
we will: 1) perform a DNase I sensitivity and hypersensitivity analysis of
the perforin gene locus; 2) analyze potential regulatory domains by reporter
gene assays; and 3) generate and analyze mice that are transgenic for
regulatory DNAs identified in the first two aims.
StatusFinished
Effective start/end date4/1/983/31/04

Funding

  • National Cancer Institute: $213,250.00
  • National Cancer Institute
  • National Cancer Institute
  • National Cancer Institute: $219,645.00

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