A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus

Boshart, Michael; Weber, Frank; Jahn, Gerhard; Dorsch-Häsler, Karoline; Fleckenstein, Bernhard; Schaffner, Walter

doi:10.1016/s0092-8674(85)80025-8

Cited by 1,091 publications

(667 citation statements)

References 69 publications

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“…These estimates are important because they serve as a baseline for interpreting the physiological e ects observed in BRCA1 transfection experiments. The CMV promoter vectors that we have used produce mRNA levels (Boshart et al, 1985;Akrigg et al, 1985) that greatly exceed the maximum prevalence of endogenous BRCA1 transcripts in non-tranfected cells. Thus, whether or not full length BRCA1 promotes cell killing in natural situations remains to be demonstrated.…”

Section: Discussionmentioning

confidence: 99%

Differential subcellular localization, expression and biological toxicity of BRCA1 and the splice variant BRCA1-Δ11b

et al. 1997

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The mechanism of BRCA1 tumor suppression in human breast and ovarian cells is the focus of intense investigation. In this report, full length BRCA1 (230 kDa) introduced into cells with CMV promoter constructs was nuclear when transgene expression was low whereas high expression resulted in cytoplasmic accumulation, aberrant nuclear and cell morphology. A nuclear localization signal (NLS) was mapped to BRCA1 amino acid positions 262 ± 570. We describe a splice variant, BRCA1-D11b, missing the majority of exon 11 including the NLS. Exogenous BRCA1-D11b (110 kDa) was cytoplasmic and, unlike the full-length protein, overexpression of the protein encoded by the variant did not appear to be toxic. RNA probe titrations and RT ± PCR demonstrated that BRCA1 and D11b transcripts are coexpressed in a wide variety of cells and tissues. Interestingly, BRCA1-D11b message was greatly reduced or absent in several breast and ovarian tumor lines relative to exon 11 transcripts and a D9,10 splice variant. Taken together our results suggest that fulllength BRCA1 and BRCA1-D11b may have distinct roles in cell growth regulation and tumorigenesis.

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Section: Discussionmentioning

confidence: 99%

Differential subcellular localization, expression and biological toxicity of BRCA1 and the splice variant BRCA1-Δ11b

et al. 1997

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“…The EphA2 receptor tyrosine kinase is a critical component in the control of cell growth and development (19). The variable light and variable heavy genes of mAb 12G3H11 were individually cloned into mammalian expression vectors encoding a human CMV major immediate early enhancer, promoter, and 5Ј-untranslated region (20). In this system, a human ␥1 chain is secreted along with a human chain (21).…”

Section: General Description Of the Test Ab And Expression Vectorsmentioning

confidence: 99%

“…Surprisingly, despite having been the object of study for over 20 years, the precise relationship between these parameters and complement activation is still unclear. For instance, conflicting results exist whether at least a portion of the hinge region is needed for complement-dependent cytotoxicity (CDC).…”

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confidence: 99%

Modulation of the Effector Functions of a Human IgG1 through Engineering of Its Hinge Region

Dall’Acqua¹,

Cook²,

Damschroder³

et al. 2006

The Journal of Immunology

124

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We report here the engineering of a humanized anti-human EphA2 mAb (mAb 12G3H11) in an effort to explore the relationship between the hinge of a human IgG1 and its effector functions. mAb 12G3H11, used here as a model, is directed against the human receptor tyrosine kinase EphA2, which is an actively investigated target for cancer therapy due to its up-regulation in many cancer cells. Various rational modifications were introduced into the hinge region of mAb 12G3H11. These mutations were predicted to modulate the hinge’s length, flexibility, and/or biochemical properties. We show that the upper and middle hinge both play important, although functionally distinct roles. In particular, middle hinge modifications predicted to decrease its rigidity or length as well as eliminating either one of its two cysteine residues had a strong negative impact on C1q binding and complement-dependent cytotoxicity. Disruption of covalent bonds between both H chains may account in part for these effects. We also describe middle hinge mutants with a significantly decreased ability to bind FcγRIIIA and trigger Ab-dependent cell-mediated cytotoxicity. Conversely, we also generated upper hinge mutants exhibiting an increase in C1q binding and complement-dependent cytotoxicity activity. Therefore, this approach represents a novel strategy to fine-tune the biological activity of a given human IgG1. We also define, for the first time in such a systematic fashion, the relationship between various characteristics of the middle and upper hinge and the corresponding effector functions.

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“…Reporter plasmids The CMV-luc reporter plasmid was constructed by subcloning of the CMV promoter region containing four putative NF-kB binding sites (nucleotides 7671 to +77, Boshardt et al, 1985) derived from pCRNCM-neo into the BglII site of pGL2 basic vector (Promega). The SV40-gal plasmid was kindly provided by Dr D Bar-Sagi, Stony Brook, NY.…”

Section: Plasmids and Constructsmentioning

confidence: 99%

A chicken c-Rel-estrogen receptor chimeric protein shows conditional nuclear localization, DNA binding, transformation and transcriptional activation

et al. 1998

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In this report, we characterize the biological and biochemical properties of a conditional protein containing chicken c-Rel fused to the hormone-binding domain of the human estrogen receptor. This chimeric c-RelER protein causes estrogen-dependent, but otherwise c-Relspeci®c, transformation of avian ®broblasts in vitro. Our results demonstrate that c-RelER heterodimerizes with wild-type c-Rel and forms speci®c complexes with IkB-a. Estrogen causes translocation of c-RelER to the nucleus and stabilizes its binding to DNA. Hormone-activated cRelER induces transcription of at least four cellular genes that are constitutively active in wild-type c-Reltransformed ®broblasts. Two distinct cell populations were examined that di ered with respect to their growth phenotypes. The growth of ®broblasts with moderate expression levels of c-RelER was stimulated by estrogen. In contrast, the addition of estrogen to cells with high cRelER expression levels resulted in inhibition of cytokinesis and the arrest of growth. The carboxy terminal transactivation domain of c-Rel was required for the induction of these e ects since neither v-Rel nor c-Rel deletion mutants were able to induce similar changes. Taken together, our results demonstrate that high levels of c-Rel expression can a ect cell cycle control and/or cytokinesis. Furthermore, they also indicate that the biological properties of c-Rel in cell growth and di erentiation will potentially di er depending on the level of expression.

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A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus

Cited by 1,091 publications

References 69 publications

Differential subcellular localization, expression and biological toxicity of BRCA1 and the splice variant BRCA1-Δ11b

Differential subcellular localization, expression and biological toxicity of BRCA1 and the splice variant BRCA1-Δ11b

Modulation of the Effector Functions of a Human IgG1 through Engineering of Its Hinge Region

A chicken c-Rel-estrogen receptor chimeric protein shows conditional nuclear localization, DNA binding, transformation and transcriptional activation

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