Similarity between nuclear matrix proteins of various cells revealed by an improved isolation method

Gerner, Christopher; Holzmann, Klaus; Grimm, Rudolf; Sauermann, Georg

doi:10.1002/(sici)1097-4644(19981201)71:3<363::aid-jcb5>3.0.co;2-w

Cited by 49 publications

(58 citation statements)

References 28 publications

(38 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover, the high proportion in our nucleolar fractions of proteins involved in well-established nucleolar functions strongly indicates that our nucleolar fraction is highly enriched in nucleoli. Several other proteins found within nucleoli in the current analysis are involved in the ultrastructural organization of the nucleus: fibrous proteins (5.1%) and structural proteins of the chromatin (2.8%) (Verheijen et al, 1986;Gerner et al, 1998;Belmont et al, 1999;Jung et al, 2000;Bergquist et al, 2001). Five proteins (2.3%) belong to the DNA-dependent protein kinase system, a multifunctional complex shown to be involved in many biological processes such as DNA repair and telomere maintenance (Dynan and Yoo, 1998;Featherstone and Jackson, 1999;d'Adda di Fagagna et al, 2001).…”

Section: Discussionmentioning

confidence: 85%

Functional Proteomic Analysis of Human Nucleolus

Scherl

Couté

Déon

et al. 2002

MBoC

443

378

View full text Add to dashboard Cite

The notion of a “plurifunctional” nucleolus is now well established. However, molecular mechanisms underlying the biological processes occurring within this nuclear domain remain only partially understood. As a first step in elucidating these mechanisms we have carried out a proteomic analysis to draw up a list of proteins present within nucleoli of HeLa cells. This analysis allowed the identification of 213 different nucleolar proteins. This catalog complements that of the 271 proteins obtained recently by others, giving a total of ∼350 different nucleolar proteins. Functional classification of these proteins allowed outlining several biological processes taking place within nucleoli. Bioinformatic analyses permitted the assignment of hypothetical functions for 43 proteins for which no functional information is available. Notably, a role in ribosome biogenesis was proposed for 31 proteins. More generally, this functional classification reinforces the plurifunctional nature of nucleoli and provides convincing evidence that nucleoli may play a central role in the control of gene expression. Finally, this analysis supports the recent demonstration of a coupling of transcription and translation in higher eukaryotes.

show abstract

Section: Discussionmentioning

confidence: 85%

Functional Proteomic Analysis of Human Nucleolus

Scherl

Couté

Déon

et al. 2002

MBoC

443

378

View full text Add to dashboard Cite

show abstract

“…3 To investigate whether the observed NMP alterations were indeed related to ACC, we analyzed cell models for apoptosis other than Fas-treated Jurkat cells. Varying the apoptosis induction mechanisms, we treated Jurkat cells with UV-light, camptothecin or staurosporine.…”

Section: Resultsmentioning

confidence: 99%

“…3 In brief, cells were lysed in 0.05% Nonidet P-40 in hypotonic buffer (10 mM HEPES/NaOH, pH 7.4, 10 mM NaCl, 3 mM MgCl 2 ). Nuclei were pelleted at 170 g for 10 min and the resulting supernatant centrifuged at 100 000 g for 60 min to yield the cytosol.…”

Section: Isolation Of the Nuclear Matrixmentioning

confidence: 99%

See 1 more Smart Citation

Proteome analysis of nuclear matrix proteins during apoptotic chromatin condensation

et al. 2002

View full text Add to dashboard Cite

The nuclear matrix (NM) is considered a proteinaceous scaffold spatially organizing the interphase nucleus, the integrity of which is affected during apoptosis. Caspasemediated degradation of NM proteins, such as nuclear lamins, precedes apoptotic chromatin condensation (ACC). Nevertheless, other NM proteins remain unaffected, which most likely maintain a remaining nuclear structure devoid of chromatin. We, therefore, screened various types of apoptotic cells for changes of the nuclear matrix proteome during the process of apoptotic ACC. Expectedly, we observed fundamental alterations of known chromatin-associated proteins, comprising both degradation and translocation to the cytosol. Importantly, a consistent set of abundant NM proteins, some (e.g. hNMP 200) of which displaying structural features, remained unaffected during apoptosis and might therefore represent constituents of an elementary scaffold. In addition, proteins involved in DNA replication and DNA repair were found accumulated in the NM fraction before cells became irreversibly committed to ACC, a time point characterized in detail by inhibitor studies with orthovanadate. In general, protein alterations of a consistent set of NM proteins (67 of which were identified), were reproducibly detectable in Fasinduced Jurkat cells, in UV-light treated U937 cells and also in staurosporine-treated HeLa cells. Our data indicate that substantial alterations of proteins linking chromatin to an elementary nuclear protein scaffold might play an intriguing role for the process of ACC.

show abstract

“…This was followed by a nuclear matrix preparation, as has been described in detail (Gerner et al, 1998).…”

Section: Two-dimensional Gel Electrophoresis Immunoblotting and Idenmentioning

confidence: 99%

Inherent growth advantage of (pre)malignant hepatocytes associated with nuclear translocation of pro-transforming growth factor α

et al. 2004

View full text Add to dashboard Cite

The pro-peptide of transforming growth factor a (proTGFa) was recently found in hepatocyte nuclei preparing for DNA replication, which suggests a role of nuclear proTGFa for mitogenic signalling. This study investigates whether the nuclear occurrence of the propeptide is involved in the altered growth regulation of (pre)malignant hepatocytes. In human hepatocarcinogenesis, the incidence of proTGFa-positive and replicating nuclei gradually increased from normal liver, to dysplastic nodules, to hepatocellular carcinoma. ProTGFa-positive nuclei almost always were in DNA synthesis. Also, in rat hepatocarcinogenesis, proTGFa-positive nuclei occurred in (pre)malignant hepatocytes at significantly higher incidences than in unaltered hepatocytes. For functional studies unaltered (GSTp À ) and premalignant (GSTp þ ) rat hepatocytes were isolated by collagenase perfusion and cultivated. Again, DNA synthesis occurred almost exclusively in proTGFa-positive nuclei. GSTp þ hepatocytes showed an B3-fold higher frequency of proTGFa-positive nuclei and DNA replication than GSTp À cells. Treatment of cultures with the mitogen cyproterone acetate (CPA) elevated the incidence of proTGFa-positive nuclei and DNA synthesis in parallel. Conversely, transforming growth factor b1 (TGFb1) lowered both. These effects of CPA and TGFb1 were significantly more pronounced in GSTp þ than in GSTp À hepatocytes. In conclusion, nuclear translocation of proTGFa increases in the course of hepatocarcinogenesis and appears to be involved in the inherent growth advantage of (pre)malignant hepatocytes.

show abstract

Similarity between nuclear matrix proteins of various cells revealed by an improved isolation method

Cited by 49 publications

References 28 publications

Functional Proteomic Analysis of Human Nucleolus

Functional Proteomic Analysis of Human Nucleolus

Proteome analysis of nuclear matrix proteins during apoptotic chromatin condensation

Inherent growth advantage of (pre)malignant hepatocytes associated with nuclear translocation of pro-transforming growth factor α

Contact Info

Product

Resources

About