Expression pattern of a hematopoietic proteoglycan core protein gene during human hematopoiesis

Stellrecht, Christine M.; Mars, Wendy M.; Miwa, Hiroshi; Beran, Miloslav; Saunders, Grady F.

doi:10.1111/j.1432-0436.1991.tb00251.x

Cited by 35 publications

(33 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Formaldehyde gels were prepared, transferred to Hybond-N membrane (Amersham), and hybridized as described (Stellrecht et al 1993). Filters were stripped in boiling 0.1% SDS as described by the manufacturer before reprobing.…”

Section: Northern Blot Analysismentioning

confidence: 99%

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Naya¹,

Stellrecht²,

Tsai³

1995

Genes Dev.

562

449

View full text Add to dashboard Cite

The insulin gene is one of the best paradigms of tissue-specific gene expression. It is developmentally regulated and is expressed exclusively in the pancreatic 13-cell. This restricted expression is directed by a tissue-specific enhancer, within the promoter, which contains an E-box sequence. The insulin E-box binds an islet-specific protein complex, termed 3al. E-boxes bind proteins belonging to the basic helix-loop-helix (bHLH) family of transcription factors. The bHLH proteins function as potent transcriptional activators of tissue-specific genes by forming heterodimers between ubiquitous and cell-restricted family members. In addition, the cell-restricted bHLH members play an important role in specifying cell fate. To isolate the tissue-specific bHLH factor controlling insulin gene expression and study its role in islet cell differentiation, a modified yeast two-hybrid system was utilized to clone a novel bHLH factor, BETA2 ([f-cell _E-box trans-activator 2_), from a hamster insulin tumor (HIT) cell cDNA library. Northern analysis demonstrates that high-level expression of the BETA2 gene is restricted to pancreatic o~-and [~-cell lines. As expected of tissue-specific bHLH members, BETA2 binds to the insulin E-box sequence with high affinity as a heterodimer with the ubiquitous bHLH factor E47. More importantly, antibody supershift experiments clearly show that BETA2 is a component of the native insulin E-box-binding complex. Transient transfection assays demonstrate that the BETA2/E47 heterodimer synergistically interacts with a neighboring 13-cell-specific complex to activate an insulin enhancer. In contrast, other bHLH factors such as MyoD and E47, which can bind to the insulin E-box with high affinity, fail to do so. Thus, a unique, cooperative interaction is the basis by which the insulin E-box enhancer discriminates between various bHLH factors to achieve tissue-specific activation of the insulin gene.

show abstract

Section: Northern Blot Analysismentioning

confidence: 99%

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Naya¹,

Stellrecht²,

Tsai³

1995

Genes Dev.

562

449

View full text Add to dashboard Cite

show abstract

“…There is some evidence that the transfection of cells by cationic lipids is mediated by membrane-sulphated proteoglycans [25] which are expressed at different levels on haematopoietic cells depending on their lineage and differentiation state [37,38]. These observations were further investigated and it was found that proteoglycans played only a minor role in PS-ODN uptake (accounting for 10-20% of the uptake, Fig.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

2003

View full text Add to dashboard Cite

Antisense oligodeoxyribonucleotides (ODNs) can specifically inhibit gene expression, but their application to fresh human CD8 þ T cells is limited by poor spontaneous uptake (<2%). We have examined and optimized the uptake of phosphorothioate-modified oligodeoxyribonucleotides (PS-ODNs) into these cells in an ex vivo expansion model. Optimal antisense treatments were found to be, for fresh CD8 þ T cells, 1 mM PS-ODNs complexed with lipofectin (LF), which resulted in 35% uptake and 10 mM PS-ODNs in the absence of LF, for cultured cells, which resulted in 95% uptake. The delivered antisenses were functional, as determined by the inhibition of protein expression. In this respect, partially phosphorothioate-modified ODNs (PS-ODNs-P) were twice as effective as completely modified (PS-ODNs-C), and the antisense specific for the cap site showed the highest protein suppression of those tested (68%). Uptake mechanisms were also investigated. To our knowledge, this is the first optimization of the delivery of antisense oligonucleotides into human CD8 þ T cells. This protocol could be used to study the function of a particular gene in cytotoxic T lymphocytes and also by those looking for a method to deliver short interfering RNA into cell lines to specifically suppress a gene of interest.

show abstract

“…For example, megakaryocytic differentiation is associated with increased serglycin expression (42,43), whereas treatment of leukemic cell lines with PMA decreased the expression of serglycin mRNA (44). During myeloblast differentiation, increased serglycin expression was shown to coincide with granule biogenesis (45), whereas, in contrast, serglycin expression is downregulated during promyelocyte differentiation into mature neutrophils (23).…”

Section: Regulation Of Serglycin Expressionmentioning

confidence: 99%

Serglycin: A Structural and Functional Chameleon with Wide Impact on Immune Cells

Kolset

Pejler

2011

The Journal of Immunology

136

130

View full text Add to dashboard Cite

Among the different proteoglycans expressed by mammals, serglycin is in most immune cells the dominating species. A unique property of serglycin is its ability to adopt highly divergent structures, because of glycosylation with variable types of glycosaminoglycans when expressed by different cell types. Recent studies of serglycin-deficient animals have revealed crucial functions for serglycin in a diverse array of immunological processes. However, its exact function varies to a large extent depending on the cellular context of serglycin expression. Based on these findings, serglycin is emerging as a structural and functional chameleon, with radically different properties depending on its exact cellular and immunological context.

show abstract

Expression pattern of a hematopoietic proteoglycan core protein gene during human hematopoiesis

Cited by 35 publications

References 36 publications

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

Serglycin: A Structural and Functional Chameleon with Wide Impact on Immune Cells

Contact Info

Product

Resources

About

Expression pattern of a hematopoietic proteoglycan core protein gene during human hematopoiesis

Cited by 35 publications

References 36 publications

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor.

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8+ T‐Cell Ex Vivo Expansion Model

Serglycin: A Structural and Functional Chameleon with Wide Impact on Immune Cells

Contact Info

Product

Resources

About

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model