The Linked Human Elongation Initiation Factor 4A1 (EIF4A1) and CD68 Genes Map to Chromosome 17p13

Jones, Emma; Quinn, Carmel M.; See, Chee Gee; Montgomery, Douglas S.; Ford, M. J.; Kölble, Konrad; Gordon, Siamon; Greaves, David R.

doi:10.1006/geno.1998.5515

Cited by 12 publications

(9 citation statements)

References 4 publications

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“…A 2940-bp BstXI fragment was purified from cosmid CD68C1 (10). The BstXI fragment was rendered blunt ended by incubation with T4 DNA polymerase and cloned into the EcoRV site of pBluescript SK Ϫ (Stratagene, La Jolla, CA) to give plasmid pCD68Bst3-2.…”

Section: Construction Of Promoter Reporter Plasmidsmentioning

confidence: 99%

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Multiple Ets Factors and Interferon Regulatory Factor-4 Modulate CD68 Expression in a Cell Type-specific Manner

O’Reilly

Quinn

El‐Shanawany

et al. 2003

Journal of Biological Chemistry

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CD68 is a transmembrane glycoprotein expressed in all cells of the mononuclear phagocyte lineage including monocytes and tissue resident macrophages. Deletion analysis of the 5-flanking sequences of the gene demonstrated that the proximal ؊150-bp sequence of the CD68 promoter exhibits high level promoter activity in macrophages. Mutations that abolish Ets factor binding at positions ؊106 and ؊89 reduce promoter activity in macrophages to 12 and 30%, respectively. Band shift experiments show that PU.1 associates with the ؊89 site whereas, Elf-1 preferentially binds the ؊106 Ets binding site and enhances CD68 activity in vitro. Furthermore, chromatin immunoprecipitation experiments confirm that Elf-1 and PU.1 associate with the CD68 proximal promoter in vivo in THP-1 cells. PU.1 does not bind to the CD68 promoter alone but instead forms heterocomplexes with members of the interferon regulatory factor family (IRF) including IRF-4 and IRF-8. IRF-4 and IRF-8 typically mediate transcriptional activation when associated with PU.1 on composite elements. However, our data show that PU.1/IRF-4 and IRF-8 heterocomplexes down-regulate CD68 promoter activity in macrophages and repression is dependent on the integrity of both the IRF and PU.1 half-sites of this composite element. Chromatin immunoprecipitation data reveal that neither IRF-4 nor IRF-8 associate with the CD68 proximal promoter in macrophages in vivo but IRF-4 is associated with the promoter in B lymphocytes. We propose that expression of CD68 in myeloid cells requires the Ets transcription factors Elf-1 and PU.1 and CD68 expression is down-regulated in lymphoid cells by combinatorial interactions between PU.1 and IRF-4.

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Section: Construction Of Promoter Reporter Plasmidsmentioning

confidence: 99%

“…CD68 is one of 9 genes in a 140-kb region of the human chromosome 17p13 (10). It lies 669 bp 3Ј to the human eukaryotic initiation factor 4AI gene and 5Ј to a MPDU1 gene, which encodes a repressor of the Lec 15 and Lec 35 glycosylation mutants of Chinese hamster ovary cells.…”

mentioning

confidence: 99%

Multiple Ets Factors and Interferon Regulatory Factor-4 Modulate CD68 Expression in a Cell Type-specific Manner

O’Reilly

Quinn

El‐Shanawany

et al. 2003

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

“…These differences may be the result of the unusual genomic location of the CD68 gene locus, with the ATG initiation codon of the CD68 gene lying 670 bp downstream of the ubiquitously expressed eukaryotic Initiation Factor 4AI (eIF4AI) gene on chromosome 17p13. 35 This means that the genetic elements required to restrict gene expression to ms are likely to be condensed in the intergenic region, while the 3Ј end of the eIF4AI gene, which makes up most of the CD68L-promoter sequence, may contain negative regulatory sequences for highlevel expression in ms.The high-efficiency retroviral transduction of HSCs in this study was achieved without the ex vivo selection of transduced cells. Several reports have suggested that preselection of transduced HSCs, often through FACS sorting of EGFP ϩ cells, is a prerequisite for generating long-lasting gene expression in vivo.…”

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confidence: 99%

Gene therapy of apolipoprotein E–deficient mice using a novel macrophage-specific retroviral vector

Gough

Raines

2003

Blood

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The use of retroviral gene transfer into hematopoietic stem cells for human gene therapy has been hampered by the absence of retroviral vectors that can generate long-lasting, lineage-specific gene expression. We developed self-inactivating retroviral vectors that incorporate generegulatory elements from the macrophage-restricted human CD68 gene. Through the transplantation of transduced murine hematopoietic stem cells (HSCs), we show that a vector incorporating a 342-base pair (bp) fragment of 5 flanking sequence from the CD68 gene, in addition to the CD68 first intron, was able to direct macrophage-specific expression of an enhanced green fluorescent protein (EGFP) reporter gene in inflammatory cell exudates and lymphoid organs in vivo. Levels of EGFP expression generated by this vector were greater than those generated by a standard Moloney murine leukemia retroviral vector, and they were stable for at least a year after transplantation of transduced HSCs. To evaluate the ability of this vector to generate therapeutically useful levels of gene expression, we transplanted apolipoprotein E (ApoE)-deficient HSCs transduced with a virus encoding ApoE into ApoEdeficient mice. Macrophages from these mice expressed levels of ApoE that were comparable to those from wild-type mice, and vector-driven expression of ApoE in macrophages was sufficient to reverse both hypercholesterolemia and atherosclerotic lesion development. The future application of this retroviral vector should provide a powerful tool to further elucidate macrophage function and for human gene therapy. IntroductionStable gene transfer into hematopoietic stem cells (HSCs) has great potential for treating inherited and acquired human diseases, as shown by the recent success in treating 2 patients with severe combined immunodeficiency. 1,2 Such an approach requires the efficient transduction of HSCs with a vector that is capable of generating long-term, high-level gene expression. At present, retroviral (including oncoretroviral and lentiviral) vectors are the only option for meeting these criteria. 3 However, expression generated by promoter elements in the viral long terminal repeat (LTR) of these vectors is often only short-lived in vivo, a problem that has hampered the clinical application of retroviral gene therapy. [4][5][6] In addition, these vectors direct expression in all progeny lineages derived from transduced HSCs, which may be detrimental in many circumstances. Thus, much work has been performed with the aim of developing vectors that can generate long-lasting transgene expression in specific hematopoietic lineages.Attempts to create oncoretroviral vectors (RVs) for lineagespecific or cell type-specific gene expression have focused on the incorporation of lineage-specific promoter elements into RVs. Work on generating RVs for targeting ␤-globin expression to red blood cells has exemplified the problems with such an approach. The insertion of an additional promoter into an RV often results in "promoter interference," reducing the activity o...

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“…The two eIF4A genes are differentially regulated in the mouse, with eIF4A I mRNA being expressed at higher levels than the eIF4A II mRNA in growing cells and with the eIF4A II mRNA being preferentially expressed in nondividing cells (Williams-Hill et al, 1997). In the human genome, the homologous EIF4A1 and EIF4A2 genes have been mapped to chromosomes 17p13 and 18p11.2, respectively (Jones et al, 1998;Sudo et al, 1995).…”

Section: Introductionmentioning

confidence: 98%

The Human Eukaryotic Initiation Factor 4AI Gene (EIF4A1) Contains Multiple Regulatory Elements That Direct High-Level Reporter Gene Expression in Mammalian Cell Lines

et al. 1999

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The Linked Human Elongation Initiation Factor 4A1 (EIF4A1) and CD68 Genes Map to Chromosome 17p13

Cited by 12 publications

References 4 publications

Multiple Ets Factors and Interferon Regulatory Factor-4 Modulate CD68 Expression in a Cell Type-specific Manner

Multiple Ets Factors and Interferon Regulatory Factor-4 Modulate CD68 Expression in a Cell Type-specific Manner

Gene therapy of apolipoprotein E–deficient mice using a novel macrophage-specific retroviral vector

The Human Eukaryotic Initiation Factor 4AI Gene (EIF4A1) Contains Multiple Regulatory Elements That Direct High-Level Reporter Gene Expression in Mammalian Cell Lines

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