RNA sequences specifically associated with mouse intracisternal A particles

Lueders, Kira K.; Segal, Shoshana; Kuff, Edward L.

doi:10.1016/0092-8674(77)90319-1

Cited by 48 publications

(67 citation statements)

References 32 publications

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“…The RNA pellets were washed with cold 80% ethanol and stored in moist form at -28°C. Cytoplasmic RNA was prepared from MOPC-104E tumor tissue by an SDS-phenol procedure (30).…”

Section: Methodsmentioning

confidence: 99%

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Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability.

Kuff¹,

Fewell²

1985

Mol. Cell. Biol.

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Intracisternal A-particle (IAP)-specific sequences were 5-to 10-fold enriched in polyadenylated RNA from BALB/cJ thymus as compared with RNAs from liver, spleen, and kidney. The major transcripts of 7.2 and 5.4 kilobases were the same size as those found in an TAP-rich neuroblastoma cell line. The absolute levels and proportions of these transcripts varied in thymuses from mice of different inbred strains. With antiserum prepared against p73, the main IAP structural protein, several size classes of IAP-related proteins were immunoprecipitated from extracts of thymus cells incubated with [35S]methionine; these included p73 itself and a group of polypeptides in the size range of 114 to 120 kilodaltons (pll4-pl20). The inbred strains showed marked characteristic differences in the electrophoretic patterns of their IAP-related proteins. Earlier studies showed that the 7.2-kilobase RNA from neuroblastoma IAPs coded for p73 in a cell-free translation system. Correlations between the RNA and protein patterns in thymuses of the different inbred strains indicated that 5.4-kilobase RNA gives rise to the pll4-pl20 polypeptides. Metabolically labeled p120 was found to include methionine-containing tryptic peptides of p73 plus additional peptides consistent with its larger size. In vivo labeling kinetics showed that the pll4-pl20 polypeptides were not major precursors of p73 in intact neuroblastoma cells. This study shows that IAP gene expression in mouse thymus is genetically determined and that a novel class of IAP-related polypeptides can be expressed independently of the major particle structural protein.Intracisternal A-particles (IAPs) are retrovirus-like entities found in many types of mouse tumor cells (20,45). They contain a group-specific internal structural protein, p73 (20,32), and a DNA polymerase activity with properties of reverse transcriptase (42, 43). The isolated particles also contain specific polyadenylated [poly(A)] RNAs (30) ranging in size from 7.2 to about 3.5 kilobases (kb) (34,36,41). These RNA species, which are related to one another in sequence, vary in relative proportion in particles isolated from different tumor sources. Endogenous provirus-like elements homologous to the IAP-associated RNAs ("IAP genes") are reiterated 1,000-fold per haploid genome in Mus musculus (28). The full-size genetic unit is 7.3 kb in length and is colinear with the 7.2-kb transcript (21). Shorter elements with internal deletions are also found (21, 34), and one group of these, the so-called type IT elements (40), are known to give rise to some of the shorter IAP-specific RNAs (34, 41). The IAP genetic elements share many structural properties with integrated retroviral proviruses and certain transposable elements in Drosophila spp. and yeasts (8a). Recently, IAP genes have been found to appear in novel locations in the genome of certain IAP-rich BALB/c mouse myeloma and hybridoma cell lines and to affect the function of genes at the various target sites (5,7,9,10,18,19,41). Thus, on occasion they can act as mobile e...

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“…The RNA pellets were washed with cold 80% ethanol and stored in moist form at -28°C. Cytoplasmic RNA was prepared from MOPC-104E tumor tissue by an SDS-phenol procedure (30).…”

Section: Methodsmentioning

confidence: 99%

“…Poly(A) RNA was isolated by chromatography on oligodeoxythymidylate-cellulose (type 7, P.-L. Biochemicals) as previously described (30). All preparations were cycled twice over the column to ensure complete removal of DNA.…”

Section: Methodsmentioning

confidence: 99%

Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability.

Kuff¹,

Fewell²

1985

Mol. Cell. Biol.

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“…Using labeled IAP RNA as a probe in blot-hybridization experiments, we have identified a characteristic electrophoretic pattern of reactive fragments generated by restriction endonuclease cleavage of mouse DNA. From the genomic blots, we deduced a composite restriction map for a 6.5-to 7-kilobase (kb) DNA (1,2). These sequences were contained in polyadenylylated RNA molecules of several discrete sizes ranging from 29S to 35S (3).…”

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confidence: 99%

“…Previously, we have identified a group of RNAs characteristically associated with IAPs prepared from mouse tumor cells (1,2). These sequences were contained in polyadenylylated RNA molecules of several discrete sizes ranging from 29S to 35S (3).…”

mentioning

confidence: 99%

“…All of the RNA species contain related sequences as judged by hybridization studies (2), and both the 35S RNA from neuroblastoma A particles and the predominant 29S species from myeloma particles have been shown to direct the cell-free synthesis of the major (73,000-dalton) A-particle structural protein (3). There are no close sequence homologies between the genomic RNA of IAPs and those of representative type B and type C viruses of M. musculus (1,2,4,5). However,-IAPs do share partial sequence homology (6) with a distinctive class of.retrovirus endogenous to the Asian murine species M. cervicolor and M. caroli (7,8).…”

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confidence: 99%

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Intracisternal A-particle genes: identification in the genome of Mus musculus and comparison of multiple isolates from a mouse gene library.

Lueders¹,

Kuff²

1980

Proc. Natl. Acad. Sci. U.S.A.

113

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The genome of Mus musculus contains multiple copies (500-1000) of DNA sequences related to the 35S RNA of intracisternal type A particles (LAPs). Using labeled IAP RNA as a probe in blot-hybridization experiments, we have identified a characteristic electrophoretic pattern of reactive fragments generated by restriction endonuclease cleavage of mouse DNA. From the genomic blots, we deduced a composite restriction map for a 6.5-to 7-kilobase (kb) DNA (1,2). These sequences were contained in polyadenylylated RNA molecules of several discrete sizes ranging from 29S to 35S (3). All of the RNA species contain related sequences as judged by hybridization studies (2), and both the 35S RNA from neuroblastoma A particles and the predominant 29S species from myeloma particles have been shown to direct the cell-free synthesis of the major (73,000-dalton) A-particle structural protein (3). There are no close sequence homologies between the genomic RNA of IAPs and those of representative type B and type C viruses of M. musculus (1, 2, 4, 5). However,-IAPs do share partial sequence homology (6) with a distinctive class of.retrovirus endogenous to the Asian murine species M. cervicolor and M. caroli (7,8).Hybridization kinetics have indicated that DNA sequences homologous to IAP RNAs are reiterated to the extent of 500 or more copies per haploid genome of both somatic and germ cells of M. musculus (6, 9). On the other hand, only'20-25 copies of the related genes were found in the DNAs of M. cervicolor and M. caroli (6-8, 10), from whose progenitors M. musculus diverged about 5 million years ago (7, 11). Thus, the evolution of The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 3571M. musculus appears to have been accompanied by a striking amplification of IAP-related genetic information, which now makes up about 0.3% of the total cellular DNA.In situ hybridization studies show that these sequences are distributed among many, perhaps all, of the mouse chromosomes (9). Beyond this, there has been no information about the internal structure of the TAP genes, their arrangement within the mouse genome, and the degree of variation among individual members of this extensive genetic family. This report describes results of studies addressed to these questions.MATERIALS AND METHODS Blot Hybridization. High molecular weight DNA was prepared (9) from BALB/c mouse myeloma MOPC-104E, a neuroblastoma tissue culture line N4 of A/Jax origin, livers of NIH Swiss mice, and the F9 teratoma cell line derived from a tumor in strain 129 mice (12). DNAs were digested to completion with restriction endonucleases (New England BioLabs) and the digests were electrophoresed in agarose gels. DNA fragments were recovered from the gels by binding on glass microbeads (13) or were transferred to cellulose nitrate sheets by a modification (14) of the Southern blot technique (15). For use a...

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Restriction enzyme evidence for Alu sequence‐mediated dispersion of microinjected genes in transgenic mice

Rubinstein¹,

Gordon²

1987

Dev. Genet.

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A human bacteriophage clone containing adult beta-globin genes with four Alu sequences was microinjected to produce transgenic mice. Southern blot analysis on the spleen of a transgenic mouse revealed an unusual hybridization pattern that suggested extensive dispersion of human DNA throughout the mouse genome. This pattern was reproducible using several restriction enzymes, including a noncutting enzyme. The hybridization pattern was not observed in other tissues, and sequences were not detected in progeny using the bacteriophage probe. However, hybridization of spleen DNA of offspring against a human Alu probe revealed genetic transmission of human Alu sequences. The results suggest dispersion of microinjected Alu sequences throughout the genome.

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RNA sequences specifically associated with mouse intracisternal A particles

Cited by 48 publications

References 32 publications

Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability.

Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability.

Intracisternal A-particle genes: identification in the genome of Mus musculus and comparison of multiple isolates from a mouse gene library.

Restriction enzyme evidence for Alu sequence‐mediated dispersion of microinjected genes in transgenic mice

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