p105, the NF-kappa B p50 precursor protein, is one of the cellular proteins complexed with the v-Rel oncoprotein in transformed chicken spleen cells.

The v-Rel oncoprotein of the avian Rev-T retrovirus is a member of the Rel/NF-kappa B family of transcription factors. The mechanism by which v-Rel malignantly transforms chicken spleen cells is not precisely known. To gain a better understanding of functions needed for transformation by v-Rel, we have now characterized the activities of mutant v-Rel proteins that are defective for specific protein-protein interactions. Mutant v-delta NLS, which has a deletion of the primary v-Rel nuclear localizing sequence, does not interact efficiently with I kappa B-alpha but still transforms chicken spleen cells approximately as well as wild-type v-Rel, indicating that interaction with I kappa B-alpha is not essential for the v-Rel transforming function. A second v-Rel mutant, v-SPW, has been shown to be defective for the formation of homodimers, DNA binding, and transformation. However, we now find that v-SPW can form functional DNA-binding heterodimers in vitro and in vivo with the cellular protein NF-kappa B p-52. Most strikingly, coexpression of v-SPW and p52 from a retroviral vector can induce the malignant transformation of chicken spleen cells, whereas expression of either protein alone cannot. Our results are most consistent with a model wherein Rel homodimers or heterodimers must bind DNA and alter gene expression in order to transform lymphoid cells.

Download Full-text

NF-kappa B p100 is one of the high-molecular-weight proteins complexed with the v-Rel oncoprotein in transformed chicken spleen cells.

Journal of Virology ◽

10.1128/jvi.67.12.7612-7617.1993 ◽

1993 ◽

Vol 67 (12) ◽

pp. 7612-7617 ◽

Cited By ~ 15

Author(s):

S Sif ◽

T D Gilmore

Keyword(s):

Molecular Weight ◽

High Molecular Weight ◽

Spleen Cells ◽

Nf Kappa B ◽

Chicken Spleen ◽

High Molecular Weight Proteins

Download Full-text

Development of Eimeria tenella in MDBK cell culture with a note on enhancing effect of preincubation with chicken spleen cells

The Korean Journal of Parasitology ◽

10.3347/kjp.1989.27.2.87 ◽

1989 ◽

Vol 27 (2) ◽

pp. 87 ◽

Cited By ~ 3

Author(s):

J Y Chai ◽

S H Lee ◽

W H Kim ◽

C K Yun

Keyword(s):

Cell Culture ◽

Eimeria Tenella ◽

Mdbk Cell ◽

Spleen Cells ◽

Enhancing Effect ◽

Chicken Spleen

Download Full-text

An Activating Mutation (Ser525Pro) within the Transactivation Domain of REL in Two Patients with Human B-Cell Lymphomas Enhances REL’s In Vitro Transforming Activity.

Blood ◽

10.1182/blood.v106.11.2617.2617 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2617-2617

Author(s):

Heiko Trautmann ◽

Daniel T. Starczynowski ◽

Christiane Pott ◽

Lana Harder ◽

Norbert Arnold ◽

...

Keyword(s):

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Transactivation Domain ◽

Wild Type ◽

Spleen Cells ◽

B Cell Lymphomas ◽

Chicken Spleen ◽

Human B Cell

Abstract REL/NF-κB transcription factors are implicated in the control of apoptosis and cell growth particular in hematopoetic lineages. The REL locus at chromosomal region 2p13–16 is frequently amplified in B-cell lymphomas including diffuse-large B-cell lymphoma (DLBCL) and may play a role in lymphomagenesis. Overexpression of wild-type REL can transform chicken lymphoid cells in culture, and several experimentally-generated mutations within the REL C-terminal transactivation domain (TAD) have been previously shown to enhance REL’s transforming ability. We analysed 83 B-cell lymphomas included in the ‘Deutsche Krebshilfe’ funded network „Molecular Mechanisms in Malignant Lymphoma“ for the presence of activating mutations in the coding region of REL. We performed a systematic dHPLC screening for mutation discovery and identified an identical point mutation in two human B-cell lymphomas (a t(14;18)-positive follicular lymphoma and a mediastinal B-cell lymphoma) that changes Ser525 to Pro within the REL TAD. In the mediastinal B-cell lymphoma, the mutation in REL was proven to be of germline origin. FISH showed an amplification of the REL locus in the tumor cells of this case. Quantitative allelic discrimination of S525P indicates that the mutant REL gene was over-represented in both cases. By in vitro experiments we could show that the S525P mutation enhances the in vitro transforming ability of REL in chicken spleen cells. In addition, REL-S525P differs from wild-type REL in its ability to activate certain κB site-containing reporter plasmids in transient transfection assays. In particular, REL-S525P has a reduced ability to activate the human manganese superoxide dismutase (MnSOD) promoter in A293 cells; however, the MnSOD protein is over-expressed in REL-S525P-transformed chicken spleen cells as compared to wild-type REL-transformed cells. Ser525 of REL falls within a sequence that is similar to other known phosphorylation sites of the IκB kinase, and REL-S525P shows a reduced ability to be phosphorylated by IKKα in vitro. The S525P mutation reduces IKKα- and TNFα-stimulated transactivation by REL, as measured in GAL4 reporter assays. Furthermore, REL-S525P-transformed chicken spleen cells are more resistant to TNFα-induced cell death than cells transformed by wild-type REL. These results represent the first identification of a tumor-derived activating mutation in the REL proto-oncogene, and they suggest that the S525P mutation contributes to the development of human B-cell lymphomas by altering REL’s ability to induce target gene expression by affecting an IKKα-regulated transactivation activity.

Download Full-text

Molecular hybridization of RNA from chicken spleen cells after immunization

Cellular and Molecular Life Sciences ◽

10.1007/bf01896920 ◽

1970 ◽

Vol 26 (4) ◽

pp. 411-412

Author(s):

St. Stipek ◽

K. Raska ◽

J. Ivanyi

Keyword(s):

Molecular Hybridization ◽

Spleen Cells ◽

Chicken Spleen

Download Full-text

INOSITOL AND THE PROTECTION OF CELLS: II. THE RELEASE OF NUCLEIC ACIDS FROM NORMAL AND MALIGNANT CELLS

Canadian Journal of Biochemistry ◽

10.1139/o64-020 ◽

1964 ◽

Vol 42 (2) ◽

pp. 167-177 ◽

Cited By ~ 5

Author(s):

R. Bather ◽

Irene Sebastian ◽

S. J. Webb

Keyword(s):

Base Composition ◽

Cell Types ◽

Prolonged Treatment ◽

Spleen Cells ◽

Ehrlich Cells ◽

Chicken Cell ◽

Chicken Spleen ◽

Ehrlich Ascites ◽

Ehrlich Ascites Cells ◽

Embryo Cells

Various normal mouse or chicken cell types or Ehrlich ascites cells were incubated for periods of up to 3 hours in either Hanks' balanced salt solution (BSS) or 5% inositol solution. Cells incubated in inositol invariably showed better viability as judged by erythrosin staining than those incubated in BSS. Ehrlich cells incubated in BSS showed a reduced ability to take up oxygen after 90 minutes incubation while those in inositol did not. Mouse embryo and chick embryo cells were able to grow in tissue culture even after prolonged treatment with 5% inositol.Maintenance of cell viability was accompanied by a greater loss of RNA from the cell in inositol compared with BSS. This loss appeared to be relatively greater from normal cells than from Ehrlich cells, and in the case of chicken spleen the loss of RNA after 2 hours incubation in inositol was 10 or more times greater than in BSS. Purification and base composition data are presented for inositol-extracted RNA from normal chicken spleen cells, Ehrlich cells, and a subline of Ehrlich. Differences in base composition are apparent for each cell type. Some implications of these findings are discussed.

Download Full-text

Identification of a rel-related protein in the nucleus during the S phase of the cell cycle.

Molecular and Cellular Biology ◽

10.1128/mcb.13.10.6147 ◽

1993 ◽

Vol 13 (10) ◽

pp. 6147-6156 ◽

Cited By ~ 5

Author(s):

R B Evans ◽

P D Gottlieb ◽

H R Bose

Keyword(s):

Cell Cycle ◽

Amino Acids ◽

S Phase ◽

Cellular Proteins ◽

Related Protein ◽

Carboxy Terminus ◽

Nf Kappa B ◽

Western Immunoblot ◽

Molecular Masses ◽

Carboxy Terminal

The c-rel proto-oncogene encodes a 75-kDa protein (p75c-rel) which is present in the cytosol of chick embryo fibroblasts (CEF) associated with a distinct set of cellular proteins with molecular masses of 40, 115, and 124 kDa. CEF cultures arrested in S phase of the cell cycle, or enriched for G2 or mitotic cells, were examined to determine whether the expression of c-rel was altered during the cell cycle. Levels of p75c-rel remained constant in all portions of the cell cycle examined; however, a Rel-related protein with an apparent molecular mass of 64 kDa was detected in nuclei of S-phase cells. As cells enter G2, the level of this protein in the nucleus decreases. This protein reacts with antiserum generated against the carboxy terminus of p75c-rel in radioimmunoprecipitations and Western immunoblot experiments and was also detected in a Western immunoblot with antiserum generated against the first 161 amino acids of pp59v-rel. Thus, unlike other Rel/NF-kappa B family members, p64 has carboxy-terminal homology with c-Rel. The majority of peptides generated by partial proteolytic cleavage of p64 are shared with peptides generated by digestion of p75c-rel and/or pp59v-rel. We suggest that this protein represents a new member of the Rel family of transcription factors and is located in the nucleus of avian fibroblasts during S phase of the cell cycle.

Download Full-text