Differential Distribution of Heterogeneous Nuclear Ribonucleoproteins in Rat Tissues

Faura, M.; Renau‐Piqueras, Jaime; Bachs, Oriol; Bosser, Ramon

doi:10.1006/bbrc.1995.2811

Cited by 14 publications

(4 citation statements)

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“…All tissues from wild type mice expressed a single, 1.8 kb transcript, consistent with previous studies [36,37]. Mice homozygous for the 1B4 provirus expressed the 1.8 kb transcript as well as an additional, larger transcript (Figure 4).…”

Section: Resultssupporting

confidence: 89%

Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene

2003

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BackgroundTagged sequence mutagenesis is a process for constructing libraries of sequenced insertion mutations in embryonic stem cells that can be transmitted into the mouse germline. To better predict the functional consequences of gene entrapment on cellular gene expression, the present study characterized the effects of a U3Neo gene trap retrovirus inserted into an intron of the hnRNP A2/B1 gene. The mutation was selected for analysis because it occurred in a highly expressed gene and yet did not produce obvious phenotypes following germline transmission.ResultsSequences flanking the integrated gene trap vector in 1B4 cells were used to isolate a full-length cDNA whose predicted amino acid sequence is identical to the human A2 protein at all but one of 341 amino acid residues. hnRNP A2/B1 transcripts extending into the provirus utilize a cryptic 3' splice site located 28 nucleotides downstream of the neomycin phosphotransferase start codon. The inserted Neo sequence and proviral poly(A) site function as an 3' terminal exon that is utilized to produce hnRNP A2/B1-Neo fusion transcripts, or skipped to produce wild-type hnRNP A2/B1 transcripts. This results in only a modest disruption of hnRNPA2/B1 gene expression.ConclusionsExpression of the occupied hnRNP A2/B1 gene and utilization of the viral poly(A) site are consistent with an exon definition model of pre-mRNA splicing. These results reveal a mechanism by which U3 gene trap vectors can be expressed without disrupting cellular gene expression, thus suggesting ways to improve these vectors for gene trap mutagenesis.

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Section: Resultssupporting

confidence: 89%

Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene

2003

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“…In 1995, Kamma et al [32] and Faura et al [33] reported a cell-type-specific expression of hnRNP proteins in whole nuclei. Our present observation on tissue-dependent variations of hnRNP H and hnRNP H' contents in nuclear matrices indicates a dynamic function of these proteins.…”

Section: Discussionmentioning

confidence: 99%

Identification of Human Common Nuclear‐Matrix Proteins as Heterogeneous Nuclear Ribonucleoproteins H and H′ by Sequencing and Mass Spectrometry

Holzmann

Korosec

Gerner

et al. 1997

European Journal of Biochemistry

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Nuclear-matrix proteins were prepared from different rat and human cells and separated by twodimensional gel electrophoresis. By computer-assisted analysis of the images, two of the proteins were identified as ubiquitously occurring (common) nuclear-matrix proteins, which appeared in tissue-dependent concentrations. The two proteins that originated from human blood mononuclear cells were analyzed further. Tryptic digests of the blotted proteins were analyzed by partial peptide sequencing and matrixassisted laser-desorption ionization-time-of-flight mass spectrometry. The two human common nuclearmatrix proteins were identified as heterogeneous nuclear ribonucleoproteins (hnRNP) H and H' or their variants. Furthermore, mass analysis revealed details on the N terminus of hnRNP H.Keywords: nuclear matrix ; common nuclear-matrix protein ; heterogeneous nuclear ribonucleoprotein ; tissue specificity ; peptide sequencing.The nuclear matrix is operationally defined as the residual nuclear structure that is yielded by sequential treatment of isolated nuclei with detergents, nucleases and buffers of high ionic strength 11 1. It is considered to represent the three-dimensional fibrillar protein structure constituting the framework of the interphase nucleus. In addition to its role in maintaining the nuclear architecture and the higher order structure of chromatin, the nuclear matrix has been reported as being involved in various nuclear activities, such as DNA replication, DNA transcription, RNA processing and steroid-hormone action. Studies on selected cell types and tissues have indicated that some nuclearmatrix proteins are cell and differentiation specific, while others are shared among various cell types [2]. Stuurman et al. [3] have presented evidence for the presence of a common set of polypeptides termed minimal matrix proteins in the nuclear matrices of mouse cells. Nakayasu and Berezney [4] have reported that eight proteins named nuclear matrins are major constituents of the internal nuclear matrix of rat liver cells. Belgrader et al. were the first to determine the sequence of a defined nuclear-matrix protein, matrin 3, by cloning and sequencing [5]. The detection of other common nuclear-matrix proteins, such as nuclear lamin B [3, 41, nucleolar protein B23 [3, 41, or the nuclear-mitotic apparatus protein [6, 71, was mainly based on immunological methods. However, the number and identity of common nuclearmatrix proteins have not been established unequivocally (reviewed in [8, 91).In systematic studies, we are investigating the nuclear-matrix proteins of various human and rat cells and tissues. ComputerCorrespondence to G. Sauermann,

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“…Plus signs (ϩ), nuclear matrix proteins also present in nonhematopoietic cells. Faura et al, 1995;Kamma et al, 1995;Holzmann et al, 1997]. Therefore, it was of interest to measure the hnRNP content in the present nuclear matrix samples.…”

Section: Hnrnps and Nuclear Laminsmentioning

confidence: 99%

Nuclear matrix proteins specific for subtypes of human hematopoietic cells

Gerner

Sauermann

1999

J. Cell. Biochem.

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Nuclear matrices were prepared from isolated subtypes of human hematopoietic cells and from cultured leukemia cells. The nuclear matrix proteins were analyzed by high-resolution two-dimensional gel electrophoresis and computer-assisted image analysis. While more than 200 protein spots were shared among the cells, about 50 distinct spots were found characteristic for individual cells or groups of related cells. This allowed to differentiate between hematopoietic cells and nonhematopoietic cells, lymphocytes and myeloid cells, monocytes, neutrophils, and promyelocytic leukemia cells. B and T lymphocytes could not be differentiated. Myeloid cells with their polymorph nuclei were characterized by the presence of 13 and by the absence of seven distinct spots, as well as by low concentrations of nuclear lamins and of heterogeneous nuclear ribonucleoproteins. Neutrophils with multilobular nuclei displayed six additional spots, while lacking 18 nuclear matrix protein spots. The nuclear matrix of proliferating cells showed three distinct spots in addition to proliferating cell nuclear antigen, increased concentrations of numatrin (B23), and heterogeneous nuclear ribonucleoproteins. The described cell-specific nuclear matrix proteins may represent new markers for hematopoietic cells.

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Differential Distribution of Heterogeneous Nuclear Ribonucleoproteins in Rat Tissues

Cited by 14 publications

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Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene

Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene

Identification of Human Common Nuclear‐Matrix Proteins as Heterogeneous Nuclear Ribonucleoproteins H and H′ by Sequencing and Mass Spectrometry

Nuclear matrix proteins specific for subtypes of human hematopoietic cells

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