The structure of the mouse lymphocyte pore-forming protein perforin

Kwon, Byoung S.; Wakulchik, Mark; Liu, Chau‐Ching; Persechini, Pedro M.; Trapani, Joseph A.; Haq, Asifa K.; Kim, Yeong; Young, John Ding‐E

doi:10.1016/s0006-291x(89)80168-8

Cited by 57 publications

(23 citation statements)

References 25 publications

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“…Insert DNA derived from clone 64 was subjected to analysis by Southern blotting, using a variety of restriction endonucleases and the PFP-6 cDNA probe. The resultant hybridization pattern was identical in each case with that observed previously with mouse genomic DNA (16), suggesting that clone 64 contained the entire coding sequence of Pfp (not shown) .…”

Section: Resultssupporting

confidence: 74%

“…A comparison of the nucleotide and derived amino acid sequences of the Pfp gene revealed identity with those predicted by cDNA cloning, as reported by two groups (13,16). The protein sequence diverges over a stretch of eight residues close to its 3' end from that reported by Lowrey et al .…”

Section: Resultssupporting

confidence: 71%

“…Sense and antisense oligonucleotides were synthesized according to the cDNA sequence for mouse PFP (16) . These were used to sequence the exons in clone 64 in their entirety and to map intron-exon boundaries (Figs.…”

Section: Resultsmentioning

confidence: 99%

“…Probes used in this study were derived from two overlapping PFP cDNA clones, designated PFP-6 and PFP-15, which together comprise the entire coding sequence for mouse PFP, including -0.2 kb of 5' untranslated (TUT) sequence, but lack most of the TUT region (16). PFP-6 lacks the TUT sequences and the first 97 by of the coding region, but extends -1.3 kb further in the 3' direction (Fig.…”

Section: Methodsmentioning

confidence: 99%

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Genomic organization of the mouse pore-forming protein (perforin) gene and localization to chromosome 10. Similarities to and differences from C9.

Trapani

Kwon

Kozak

et al. 1990

The Journal of Experimental Medicine

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The molecular mechanisms involved in the delivery of cellular injury or death by CTL and NK cells have not been elucidated (1, 2). The granule-exocytosis model of cellular cytotoxicity, the most closely investigated mechanism to date, postulates that an obligate facet of the delivery of the "lethal hit" is the directional release of cytotoxic polypeptides and other molecules (the granule contents) towards the target cell (3-5). Key molecules in this process are a cytolytic protein termed perforin (5, 6), cytolysin (3), pore-forming protein (PFP)t (4) or C9-related polypeptide (C9RP) (7), and a family of killer cell-specific, highly homologous serine esterases (SE) (8,9). PFP is postulated to intercalate the target cell membrane and compromise its osmotic stability; however, the function of SEs in the cytotoxic process is less clear. Intriguing parallels have been drawn between this mechanism and the complement cascade, including the close similarities between the immunological and physicochemical properties ofPFP and C9, the ninth component ofcomplement (7,(10)(11)(12) .Many ofthe granule polypeptides have been purified to homogeneity, and cDNA clones encoding PFP (13-16) and several of the SEs (17-25) have recently been characterized. To further analyze the role of PFP in the cytolytic process and the basis for its immunological and evolutionary relationship to C9, we have investigated the This work was supported in part by U. S.

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Section: Resultssupporting

confidence: 74%

Section: Resultssupporting

confidence: 71%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Genomic organization of the mouse pore-forming protein (perforin) gene and localization to chromosome 10. Similarities to and differences from C9.

Trapani

Kwon

Kozak

et al. 1990

The Journal of Experimental Medicine

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“…The membranes were then probed with a 32 P-labeled cDNA probe for perforin (Ref. 16; a gift from Dr. Chau Ching Liu, University of Pittsburgh School of Medicine). Prehybridization was performed for 30 min at 68°C using Perfecthyb hybridization buffer.…”

Section: Perforin Mrna Detection By Northern Blot Analysismentioning

confidence: 99%

Nitric Oxide Synthase-2 and Expression of Perforin in Uterine NK Cells

Burnett

Hunt

2000

The Journal of Immunology

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In human, mouse, and rat pregnancy, maternal NK cells accumulate and differentiate at implantation sites. These cells, termed uterine NK (uNK) cells, express NO synthase (NOS)-2 and develop cytolytic molecules such as perforin and granzymes during differentiation in situ. In this study, relationships between expression of the NOS-2 gene, uNK cell population density and tissue distribution, and synthesis of perforin were investigated. Uteri from wild-type (WT) and NOS-2−/− mice were collected at gestation days (g.d.) 8, 10, 12, 14, and 16 (n, >2/g.d.). Histochemical staining failed to reveal any differences between the population densities or tissue distributions of uNK cells in WT and NOS-2−/− uteri at any stage of gestation. By contrast, immunohistochemical staining with anti-perforin Abs demonstrated significantly fewer perforin-positive uNK cells in two uterine compartments of NOS-2−/− mice in comparison to the same compartments in WT mouse uteri. Perforin-positive uNK cells were reduced in NOS-2−/− metrial glands at g.d. 8, 10, and 12 and in decidua basalis at g.d. 12 (p < 0.05). Analysis of perforin protein by immunoblotting confirmed this observation. Northern blot hybridization studies showed that loss of perforin protein in NOS-2−/− mice was accompanied by decreased steady-state levels of perforin mRNA. These results demonstrate that migration of uNK cells into the uterus, selection of residency sites, and proliferation in situ are independent of NOS-2. By contrast, their differentiation, including transcription and translation of the cytotoxic molecule perforin, was shown to rely on normal expression of the NOS-2 gene.

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Nephritogenic T cells use granzyme C as a cytotoxic mediator

Bailey¹,

Kelly²

1997

Eur J Immunol

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Cytoplasmic granules of cytotoxic T lymphocytes contain several proteins that may be involved in cell-mediated cytotoxicity. We have previously described nephritogenic T cell clones that are cytotoxic to cultured renal proximal tubular epithelial cells (MCT). One of these clones, M52.34.1, expresses perforin, a cytotoxic mediator. We investigated the expression of other granule-associated proteases of M52.34.1. Granzymes A and B have been extensively studied in T cell-mediated cytotoxicity, and associated with tissue destruction in models of transplantation. However, the activity of other granzymes has not been as extensively investigated. We focused our studies on granzyme C. Northern blots showed very high levels of granzymes B and C mRNA expression in M52.34.1 cells 3 days following T cell activation. There was no expression of granzyme A mRNA. An antisense oligonucleotide made from the 5'-upstream region of the murine granzyme C exon 1 inhibited granzyme C mRNA expression in M52.34.1 when added at a concentration of 50 microM to the culture medium for 2 days. There was no inhibition of granzyme C mRNA expression with the sense oligonucleotide. The granzyme C antisense oligonucleotide inhibited M52.34.1 cytotoxicity to MCT at effector:target ratios of 20:1 and 40:1. M52.34.1 cells mediate inflammatory interstitial nephritis following adoptive transfer. If T cells were resuspended in 200 microM of the antisense oligonucleotide prior to subcapsular transfer, the recipient kidneys showed markedly diminished tubular cell destruction, suggesting that granzyme C can also be an important mediator of cytotoxicity in vivo.

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The structure of the mouse lymphocyte pore-forming protein perforin

Cited by 57 publications

References 25 publications

Genomic organization of the mouse pore-forming protein (perforin) gene and localization to chromosome 10. Similarities to and differences from C9.

Genomic organization of the mouse pore-forming protein (perforin) gene and localization to chromosome 10. Similarities to and differences from C9.

Nitric Oxide Synthase-2 and Expression of Perforin in Uterine NK Cells

Nephritogenic T cells use granzyme C as a cytotoxic mediator

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