Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth.

Hirschhorn, Ricky R.; Aller, Patricio; Yuan, Zhi-An; Gibson, Carolyn W.; Baserga, Renato

doi:10.1073/pnas.81.19.6004

Cited by 181 publications

(142 citation statements)

References 43 publications

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“…tsAF8 cells are G1-specific, temperature-sensitive mutants originally isolated from BHK cells (12); these cells have been extensively characterized (3,12,25 Berger (9). Plasmid DNA was isolated and purified by phenol extraction and Sepharose 2B chromatography (31,32,44).…”

Section: Methodsmentioning

confidence: 99%

“…RNA isolation and blot procedures were previously described (31). Briefly, after extraction RNA was denatured and size fractionated on 1% agarose gels containing 6.6% formaldehyde.…”

Section: Methodsmentioning

confidence: 99%

“…USA, in press), c-ras (13,27,28), and c-fos (18,29,38) and genes encoding p53 (48,60), actin (13,29,46), tubulin (46), and thymidine kinase (44). Other cell cycledependent genes have been identified as cDNA clones (17,31,43) by differential screening of cDNA libraries.…”

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confidence: 99%

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Adenovirus type 2 activates cell cycle-dependent genes that are a subset of those activated by serum.

Liu¹,

Baserga²,

Mercer³

1985

Mol. Cell. Biol.

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We have studied a panel of 10 genes and cDNA sequences that are expressed in a cell cycle-dependent manner in different types of cells from different species and that are inducible by different mitogens. These include five sequences (c-myc, 4F1, 2F1, 2A9, and KC-1) that are preferentially expressed in the early part of the G, phase, three genes (ornithine decarboxylase, p53, and c-rasHa) preferentially expressed in middle or late G1, and two genes (thymidine kinase and histone H3) preferentially expressed in the S phase of the cell cycle. We have studied the expression of these genes in nonpermissive (tsAF8) and semipermissive (Swiss 3T3) cells infected with adenovirus type 2. Under the conditions of these experiments, adenovirus type 2 infection stimulates cellular DNA synthesis in both tsAF8 and 3T3 cells. However, four of the five early GI genes (c-myc, 4F1, KC-1, and 2A9) and one of the late G, genes (c-ras) are not induced by adenovirus infection, although they are strongly induced by serum. The other sequences (2F1, ornithine decarboxylase, p53, thymidine kinase, and histone H3) are activated by both adenovirus and serum. We conclude that the cell cycle-dependent genes activated by adenovirus 2 are a subset of the cell cycle-dependent genes activated by serum. The data suggest that the mechanisms by which serum and adenovirus induce cellular DNA synthesis are not identical.Cell cycle-dependent genes are operationally defined here as genes that are preferentially expressed in a specific phase of the cell cycle. A number of animal genes have already been identified whose expression is cell cycle dependent. These include histone genes (2, 22, 57), c-myc (13, 28, 35, 49; B. Calabretta, L. Kaczmarek, W. Mars, D. Ochoa, C. W. Gibson, R. R. Hirschhom, and R. Baserga, Proc. Natl. Acad. Sci. USA, in press), c-ras (13,27,28), and c-fos (18,29,38) and genes encoding p53 (48, 60), actin (13,29,46) serum and that adenovirus infection preferentially activates genes whose expression reaches a maximum in the late G1-S phase. MATERIALS AND METHODSCell line and culture conditions. tsAF8 cells are G1-specific, temperature-sensitive mutants originally isolated from BHK cells (12); these cells have been extensively characterized (3,12,25). Monolayer cultures are routinely maintained in Dulbecco modified Eagle medium containing 10% (vol/vol) donor calf serum at the PT of 34°C. To obtain quiescent (Go) cultures, tsAF8 cells are plated at 106 cells per 100-mm plate, grown to near confluence at 34°C, rinsed twice with prewarmed Hanks salt solution, fed with medium containing 0.5% (vol/vol) donor calf serum, and incubated for an additional 48 h at 34°C. Swiss 3T3 cells were cultured and made quiescent as previously described (47).Virus propagation, purification, and infection. Ad2 was grown in HeLa suspension cultures maintained in minimum essential modified suspension medium (Flow Laboratories, Rockville, Md.) with 5% fetal calf serum. Virus was purified as described previously (63)

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Section: Methodsmentioning

confidence: 99%

“…RNA isolation and blot procedures were previously described (31). Briefly, after extraction RNA was denatured and size fractionated on 1% agarose gels containing 6.6% formaldehyde.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Adenovirus type 2 activates cell cycle-dependent genes that are a subset of those activated by serum.

Liu¹,

Baserga²,

Mercer³

1985

Mol. Cell. Biol.

Self Cite

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“…The search for these important mammalian "cell division cycle" genes (5) has been carried out in a number of laboratories using differential colony hybridization screening of cDNA libraries with radiolabelled cDNA probes that reflect the quiescent and stimulated growth states (e._&. [5][6][7][8][9][10]. These and other studies have uncovered so far some forty or so genes only a few of which share identity with characterized oncogenes (reviewed in 11).…”

Section: Introductionmentioning

confidence: 99%

A growth-responsive gene (16C8) in normal mouse fibroblasts homologous to a human collagenase inhibitor with erythroid-potentiating activity: evidence for inducible and constitutive transcripts

et al. 1986

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We present the DNA sequence of an essentially full-length cDNA clone of 16C8, a growth factor-inducible gene isolated from a mouse embryo fibroblast cDNA library. The 0.9-kb mRNA encodes an Mr 22,500 protein that has substantial homology to a human protein with the reported abilities (1) to potentiate erythroid differentiation and (2) to inhibit collagenases and other tissue metalloproteinases. The N-terminus of the predicted protein has a hydrophobic nature characteristic of secreted proteins, and two potential sites for Nlinked glycosylation are present. The cytoplasmic concentration of 16C8 mRNA is maximal in mid Gl at about 6 h after serum stimulation of quiescent fibroblasts. Northern blot analysis showed a progressive reduction in the size of the induced 16C8 transcripts with increasing time after serum stimulation. This was shown to be due to the reduction in length of the poly(A) tails. S1 analysis of the 5' portion of the mRNA revealed the presence of three different species of transcript, only one of which was inducible.

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“…Although the function of calcyclin is still unknown, several observations can be made: the calcyclin gene was induced by serum, platelet-derived growth factor, or epidermal growth factor [1,4,5]. The gene product was shown to bind both zinc and calcium [6] which indicated that it is involved in processes related to signal transduction.…”

mentioning

confidence: 99%

Functional Analysis of the Human Calcyclin Gene Promoter in a Panel of Human Melanoma Cell Lines

Groningen

Weterman

Swart

et al. 1995

Biochemical and Biophysical Research Communications

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By comparing two subsequent human tumor stages we previously described calcyclin as a new potential melanoma associated neoplastic progression marker positively linked with metastasis. In this study the calcyclin expression levels in a representative panel of human melanoma cell lines were correlated with the occurrence of DNase I hypersensitive (DH) regions and potential enhancer elements in a 6 kb genomic fragment spanning the human calcyclin gene. Examination of the chromatin structure of the transcription unit revealed no qualitative differences in DH sites within the panel of tested human melanoma cells, but especially the sequences around the transcription start site and a 1.5 kb upstream region appeared more accessible to the nuclease in frequently (BLM, MV3) as compared to poorly (530, 1F6) metastasizing cells. The genomic fragments that harbor one or more DH sites were subjected to functional analysis by luciferase reporter gene assays. Thus, an enhancer element was detected between 361 and 167 bp upstream of the transcription start site. This enhancer displayed equal activating potential (2-3 fold) both in weakly and in frequently metastasizing cells and was apparently recognized by transcription factors present in both types of human melanoma cell lines. We. conclude that, in addition to a slight amplification of the encoding gene, the elevated calcyclin mRNA levels are only reflected in a selectively increased accessibility of the chromatin structure to DNasel in metastasizing melanoma cells © 1995 Academic Preas, Inc.Calcyclin is a small calcium-binding protein (10.5 kD) belonging to the SI00 family containing two calcium-binding structures, the so called EF hands. It was identified as a differentially expressed mRNA in serum-stimulated and quiescent fibro blasts [1,2] and was originally isolated from Gr specific temperature sensitive mutants of baby Syrian hamster kidney cells [3].Although the function of calcyclin is still unknown, several observations can be made: the calcyclin gene was induced by serum, platelet-derived growth factor, or epidermal growth factor [1,4,5]. The gene product was shown to bind both zinc and calcium [6] which indicated that it is involved in processes related to signal transduction.

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Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth.

Cited by 181 publications

References 43 publications

Adenovirus type 2 activates cell cycle-dependent genes that are a subset of those activated by serum.

Adenovirus type 2 activates cell cycle-dependent genes that are a subset of those activated by serum.

A growth-responsive gene (16C8) in normal mouse fibroblasts homologous to a human collagenase inhibitor with erythroid-potentiating activity: evidence for inducible and constitutive transcripts

Functional Analysis of the Human Calcyclin Gene Promoter in a Panel of Human Melanoma Cell Lines

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