THE SYNTHESIS OF ACIDIC CHROMOSOMAL PROTEINS DURING THE CELL CYCLE OF H<scp>E</scp>L<scp>A</scp> S-3 CELLS

Stein, Gary S.; Borun, Thaddeus W.

doi:10.1083/jcb.52.2.292

Cited by 270 publications

(86 citation statements)

References 35 publications

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“…During cell development, the amount of this protein varies. For instance, biochemical (21,25,38) and cytochemical (4,5,34,36,46) data have shown that at the onset of DNA synthesis a considerable increase in nuclear nonhistone protein (NNHP) is noted. An increased NNHPDNA ratio is therefore usually found in proliferating tissue (28,301.…”

mentioning

confidence: 99%

DNA and nuclear protein measurement in isolated nuclei of human endometrium

et al. 1986

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Feulgen-DNA and nuclear light green-protein measurements have been performed in isolated nuclei of normal (nonmalignant) and malignant human endometrial homogenates.The DNA content of the GO/Gl fraction of malignant endometrium showed much overlap with that of normal endometrium, or was slightly increased. Two of the 18 carcinomas were clearly aneuploid. No correlation was found between the histological grade and the DNA content. The tumors of clinical stage I1 and higher all had a higher DNA content than that of normal endometrium.The percentage of cells present in the proliferative fraction was higher in proliferative endometrium than in secretory and postmenopausal atrophic endometrium. For malignant endometrium, percentages were found comparable to that of normal endometrium or higher. No correlation was found with the histological grade. Tumors of stage I1 and higher had intermediate values compared to those of carcinomas below stage 11.The nuclear protein/l)NA ratio of malignant endometrium completely overlapped that of normal endometrium. However, for postmenopausal women, most values of the carcinomas exceeded that of normal, atrophic, endometrium. Within the tumor population, no correlation was found with the histological grade. Higher values were found with tumors of clinical stage I1 and higher.

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confidence: 99%

DNA and nuclear protein measurement in isolated nuclei of human endometrium

et al. 1986

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“…Synchrony was determined by ['~H]thymidine (New England Nuclear, Boston, Mass., 6.7 Ci/mmol) incorporation (20) and autoradiography (see Results). Ceils used for autoradiography were labeled with 10 /xCi/ml [3H]thymidine in suspension for 10 min, washed, resuspended in RSB, and fixed on slides with ethanol:chloroform:acetic acid (6:3:1).…”

Section: Cell Synchronization and Labelingmentioning

confidence: 99%

“…The synthesis of bulk chromosomal bound proteins goes on throughout the cell cycle (19) although some small proteins may be made only in S (6). These proteins showed different profiles on sodium dodecyl sulfate (SDS) gels depending on the cell cycle and changes in the rate of synthesis before DNA synthesis (20,2). Kolodny and Gross (10) analyzed total cytoplasmic proteins synthesized during the HeLa cell cycle on one-dimensional SDS gels and reported variations in certain proteins.…”

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The synthesis of ninety proteins including actin throughout the HeLa cell cycle.

Milcarek¹,

Zahn²

1978

The Journal of Cell Biology

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Abundant cytoplasmic proteins pulse-labeled with pS]methionine at specific times throughout the HeLa cell cycle were analyzed with two-dimensional gel electrophoresis. More than 300 proteins could be resolved in this way. The frequency of appearance of label in the most abundant 90 proteins, ranging from 4% to <0.1% of the total methionine incorporated, was determined at six time points in the cell cycle. 84 of these proteins were made as a similar proportion of the total at all times during the cell cycle. A nonmuscle actin protein (spot 1) identified by molecular weight and isoelectric point represented 2-4% of the total methionine incorporated at all the time points. Only six proteins were found which varied by greater than fourfold during cell division, four appearing to represent a greater proportion of the total synthesis during the period at or immediately surrounding M (spots 31b, 44, 53, and 70d). Two appear to represent a smaller percentage of total synthesis during the early (spot 78) or the total (spot 74) G2 period.KEY WORDS 2-dimensional gels cell cycle synchrony protein synthesis 9 actinThe mammalian cell in culture grows and divides in a well regulated and timed series of events. During S phase the DNA is synthesized, during M the cell divides, and between these are two gaps: G2, post DNA synthesis, and G,, postmitosis (16). Cell surface changes occur during the cycle as do the activities of a variety of enzymes (16). Protein synthesis drops 75% at mitosis as a result of a block in initiation (5). Most RNA synthesis is shut off at mitosis except for some small molecular weight species (23, 25). The mRNA persists through mitosis, however (21, 8). The synthesis of bulk chromosomal bound proteins goes on throughout the cell cycle (19) although some small proteins may be made only in S (6). These proteins showed different profiles on sodium dodecyl sulfate (SDS) gels depending on the cell cycle and changes in the rate of synthesis before DNA synthesis (20, 2). Kolodny and Gross (10) analyzed total cytoplasmic proteins synthesized during the HeLa cell cycle on one-dimensional SDS gels and reported variations in certain proteins. More detailed analysis of proteins is made possible by the two-dimensional technique (isoelectric focusing followed by SDS electrophoresis) developed by O'Farrell (15). We analyzed proteins synthesized by cells in early and late S, G~, M, and thymidine-arrested G1 by the two-dimensional technique. We have found that, of the 90 relatively abundant cytoplasmic proteins studied by frequency analysis, only eight proteins vary by greater than fourfold during the cell cycle. Nonmuscle actin (7,17,22,24) was found to represent a relatively constant percentage of total incorporation (2-4% of total) at all times studied. MATERIALS AND METHODS Cell CultureHeLa ($3) cells were grown in suspension at 37~ in Eagle's medium (4) supplemented with 7% horse serum (Flow Laboratories, Inc., Rockville, Md.). Cell cultures were tested by Microbiological Associates, Walkersville, Md., and f...

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“…Unless indicated otherwise, cells were grown for 1 day to 60 -70% confluency and treated with various concentrations of cadein1. Cells were synchronized at G 1 /S by a modified double thymidine block (15). For fluorescence-activated cell sorter analysis, cells were trypsinized, fixed with cold 70% ethanol (Ϫ20°C), treated with 100 g/ml RNase A (Sigma), and stained with 100 g/ml propidium iodide (Sigma).…”

Section: Methodsmentioning

confidence: 99%

A New Isoquinolinium Derivative, Cadein1, Preferentially Induces Apoptosis in p53-defective Cancer Cells with Functional Mismatch Repair via a p38-dependent Pathway

Jang

Ryu

Park

et al. 2010

Journal of Biological Chemistry

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We screened a protoberberine backbone derivative library for compounds with anti-proliferative effects on p53-defective cancer cells. A compound identified from this small molecule library, cadein1 (cancer-selective death inducer 1), an isoquinolinium derivative, effectively leads to a G 2 /M delay and caspasedependent apoptosis in various carcinoma cells with nonfunctional p53. The ability of cadein1 to induce apoptosis in p53-defective colon cancer cells was tightly linked to the presence of a functional DNA mismatch repair (MMR) system, which is an important determinant in chemosensitivity. Cadein1 was very effective in MMR ؉ /p53 ؊ cells, whereas it was not effective in p53 ؉ cells regardless of the MMR status. Consistently, when the function of MMR was blocked with short hairpin RNA in SW620 (MMR ؉ /p53 ؊ ) cells, cadein1 was no longer effective in inducing apoptosis. Besides, the inhibition of p53 increased the pro-apoptotic effect of cadein1 in HEK293 (MMR ؉ /p53 ؉ ) cells, whereas it did not affect the response to cadein1 in RKO (MMR ؊ /p53 ؉ ) cells. The apoptotic effects of cadein1 depended on the activation of p38 but not on the activation of Chk2 or other stressactivated kinases in p53-defective cells. Taken together, our results show that cadein1 may have a potential to be an anticancer chemotherapeutic agent that is preferentially effective on p53-mutant colon cancer cells with functional MMR.

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THE SYNTHESIS OF ACIDIC CHROMOSOMAL PROTEINS DURING THE CELL CYCLE OF HELA S-3 CELLS

Cited by 270 publications

References 35 publications

DNA and nuclear protein measurement in isolated nuclei of human endometrium

DNA and nuclear protein measurement in isolated nuclei of human endometrium

The synthesis of ninety proteins including actin throughout the HeLa cell cycle.

A New Isoquinolinium Derivative, Cadein1, Preferentially Induces Apoptosis in p53-defective Cancer Cells with Functional Mismatch Repair via a p38-dependent Pathway

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