Chromatin Structure and Gene Activity: The Role of Nonhistone Chromosomal Protein

Cartwright, Iain L.; Abmayr, Susan M.; Fleischmann, Gerhard; Lowenhaupt, Ky; Elgin, Sarah C. R.; Keene, Michael A.; Howard, Gary C.

doi:10.3109/10409238209108709

Cited by 182 publications

(39 citation statements)

References 658 publications

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“…The correlation of tissue-specific differences in general DNase I sensitivity with gene transcription has been well-documented for several genes such as globin, ovalbumin, albumin and ct-fetoprotein [13,27]. In addition, enhanced DNase I sensitivity or hypersensitivity is observed in the 5' promoter region of active genes [35,36]. Since several factors are likely to contribute to the DNase sensitivity of chromatin, such as altered conformation of nucleosomes, binding of specific proteins or changes in DNA conformation, the reduction of band intensity as a function of DNase I concentration may not be exactly comparable for different genes.…”

Section: Discussionmentioning

confidence: 98%

“…The transcriptional activation of a gene would require the initial decondensation of a domain of chromatin containing the gene This unfolding of the higher order structure can be conveniently studied by the use of endonucleases such as DNase I [27,28,35]. The correlation of tissue-specific differences in general DNase I sensitivity with gene transcription has been well-documented for several genes such as globin, ovalbumin, albumin and ct-fetoprotein [13,27].…”

Section: Discussionmentioning

confidence: 99%

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Activation of the lamin A gene during rat liver development

et al. 1996

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We have studied the regulation of expression of the Atype lamins, which are constituents of the nuclear lamina. During rat liver development, high levels of lamin A and C mRNAs were observed in 15-day fetal fiver but were barely detectable in the adult. The chromatin conformation of the lamin A gene was sensitive to DNase I in 15-day fetal liver but became mostly insensitive in the adult. Lamin A and C proteins could be detected in fetal liver and persisted in the adult. Our evidence suggests that the lamin A gene is actively transcribed early in fiver differentiation and its activity declines considerably in adult liver.Key words." Lamin A gene; Nuclear lamina; Liver development occurs primarily at the level of transcription. This differential gene expression in developing liver provides sufficient background for the study of regulatory mechanisms of other developmentally activated genes such as the A-type lamins.In order to understand how the lamin A gene is regulated, we have initially assessed lamin A expression during rat liver development by Northern analysis of RNA and nuclear runon transcription assays. The unfolding of the chromatin structure of the gene has been studied and lamin A protein levels determined. Our evidence suggests that the lamin A gene is actively transcribed by day 15 of embryonic liver development.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Activation of the lamin A gene during rat liver development

et al. 1996

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“…Numerous differences between the nuclear conformation of transcriptionally active and inactive DNA sequences have been Identified. These differences include location In euchromatin versus heterochromatin, Increased sensitivity of active genes to digestion by nucleases, an altered nucleosomal structure of active genes, and their association with specific proteins (1,2).…”

Section: Introductionmentioning

confidence: 99%

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

1985

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ABSIRACTUsing the transcriptional modulation afforded by heat shock, we found that the association of active genes with the nuclear matrix was not dependent on their level of transcription. Heat shock genes were matrix associated both before heat shock (when transcription was relatively low), and during heat shock (when transcription was greatly Increased). Conversely, the cytoplasmic actin gene was matrix associated during normal growth conditions (when transcription was high) and during heat shock (when transcription was greatly decreased). Removal of greater than 99.7% of nascent RNA during preparation of the matrices did not affect these findings. Detailed examination of the cytoplasmic actin gene revealed that Its matrix association was apparently mediated by multiple Interactions near the 5' end of the gene.

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“…Such a chromatin structure over a eucaryotic promoter may be a necessary (but not sufficient) condition for transcription from that promoter (6,12,43). In addition, hypersensitive sites have been reported which are probably not related to promoter function (3,22,28).…”

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confidence: 99%

Barriers to nuclease Bal31 digestion across specific sites in simian virus 40 chromatin.

Scott¹,

Walter²,

Cryer³

1984

Mol. Cell. Biol.

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A portion of the nucleoprotein containing viral DNA extracted from cells infected by simian virus (SV40) is preferentially cleaved by endonucleases in a region of the genome encompassing the origin of replication and early and late promoters. To explore this nuclease-sensitive structure, we cleaved SV40 chromatin molecules with restriction enzymes and digested the exposed termini with nuclease Bal3l. Digestion proceeded only a short distance in the late direction from the MspI site, but some molecules were degraded 400 to 500 base pairs in the early direction. By comparison, BglI-cleaved chromatin was digested for only a short distance in the early direction, but some molecules were degraded 400 to 450 base pairs in the late direction. These barriers to Bal3l digestion (bracketing the BglI and the MspI sites) define the borders of the same open region in SV40 chromatin that is preferentially digested by DNase I and other endonucleases. In a portion of the SV40 chromatin, Bal3l could not digest through the nuclease-sensitive region and reached barriers after digesting only 50 to 100 base pairs from one end or the other. Chromatin molecules that contain barriers in the BglI to MspI region are physically distinct from molecules that are open in this region as evidenced by partial separation of the two populations on sucrose density gradients.Viral nucleoprotein complex extracted from cells infected by simian virus 40 (SV40) resembles cellular chromatin (1,10,15,27). A short segment of the viral genome (ca. 400 base pairs [bp] extending from the SV40 origin of replication in the late direction) is hypersensitive to endonuclease cleavage (33,(38)(39)(40) and appears, in the electron microscope, as a region which is free of nucleosomes (19,32). Recently, more detailed studies have shown that, within the nuclease-sensitive site, there are subregions with differential sensitivity to DNase 1 (7,8,31,32). Presumably, these features reflect the structural details of the nucleoprotein within this region.Nuclease-hypersensitive sites usually occur over regions which contain genetic signals. Such a chromatin structure over a eucaryotic promoter may be a necessary (but not sufficient) condition for transcription from that promoter (6,12,43). In addition, hypersensitive sites have been reported which are probably not related to promoter function (3,22,28). SV40-infected cells provide a good opportunity to investigate the details of the nuclease-sensitive chromatin structure, since SV40 chromatin is present in high concentration by comparison with other genes and can easily be separated from bulk cellular chromatin.We have developed a new approach for probing nucleoprotein structure which should be applicable to the study of chromatin structure in other minichromosomes. SV40 is cleaved with a restriction enzyme which cuts at a unique position, and the exposed termini are digested with Bal3l, an exonuclease which can degrade from either 3' or 5' ends. MATERIALS AND METHODSGrowth of cells and virus infections. African green monk...

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Chromatin Structure and Gene Activity: The Role of Nonhistone Chromosomal Protein

Cited by 182 publications

References 658 publications

Activation of the lamin A gene during rat liver development

Activation of the lamin A gene during rat liver development

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

Barriers to nuclease Bal31 digestion across specific sites in simian virus 40 chromatin.

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