scholarly journals Centromere Biology: Transcription Goes on Stage

2018 ◽  
Vol 38 (18) ◽  
Author(s):  
Carlos Perea-Resa ◽  
Michael D. Blower
Keyword(s):  
Repetitive Dna ◽  
Cell Biology ◽  
Chromosomal Dna ◽  
Daughter Cells ◽  
Centromere Function ◽  
Kinetochore Assembly ◽  
Fundamental Process ◽  
A Cell ◽  
Highly Repetitive Dna ◽  
Topological Effect

ABSTRACT Accurate chromosome segregation is a fundamental process in cell biology. During mitosis, chromosomes are segregated into daughter cells through interactions between centromeres and microtubules in the mitotic spindle. Centromere domains have evolved to nucleate formation of the kinetochore, which is essential for establishing connections between chromosomal DNA and microtubules during mitosis. Centromeres are typically formed on highly repetitive DNA that is not conserved in sequence or size among organisms and can differ substantially between individuals within the same organism. However, transcription of repetitive DNA has emerged as a highly conserved property of the centromere. Recent work has shown that both the topological effect of transcription on chromatin and the nascent noncoding RNAs contribute to multiple aspects of centromere function. In this review, we discuss the fundamental aspects of centromere transcription, i.e., its dual role in chromatin remodeling/CENP-A deposition and kinetochore assembly during mitosis, from a cell cycle perspective.

Active maize genes are unmodified and flanked by diverse classes of modified, highly repetitive DNA

Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 565-576 ◽  
Author(s):  
Jeffrey L. Bennetzen ◽  
Kathrin Schrick ◽  
Patricia S. Springer ◽  
Willis E. Brown ◽  
Phillip SanMiguel
Keyword(s):  
Repetitive Dna ◽  
Dna Sequences ◽  
Copy Number ◽  
Repetitive Sequences ◽  
Restriction Enzymes ◽  
Single Copy ◽  
Chromosomal Dna ◽  
Coding Regions ◽  
Highly Repetitive Dna ◽  
Copy Dna

We have characterized the copy number, organization, and genomic modification of DNA sequences within and flanking several maize genes. We found that highly repetitive DNA sequences were tightly linked to most of these genes. The highly repetitive sequences were not found within the coding regions but could be found within 6 kb either 3′ or 5′ to the structural genes. These highly repetitive regions were each composed of unique combinations of different short repetitive sequences. Highly repetitive DNA blocks were not interrupted by any detected single copy DNA. The 13 classes of highly repetitive DNA identified were found to vary little between diverse Zea isolates. The level of DNA methylation in and near these genes was determined by scoring the digestibility of 63 recognition/cleavage sites with restriction enzymes that were sensitive to 5-methylation of cytosines in the sequences 5′-CG-3′ and 5′-CNG-3′. All but four of these sites were digestible in chromosomal DNA. The four undigested sites were localized to extragenic DNA within or near highly repetitive DNA, while the other 59 sites were in low copy number DNAs. Pulsed field gel analysis indicated that the majority of cytosine modified tracts range from 20 to 200 kb in size. Single copy sequences hybridized to the unmodified domains, while highly repetitive sequences hybridized to the modified regions. Middle repetitive sequences were found in both domains.Key words: genome organization, interspersed repetitive DNA, DNA modification.

The locomotory behaviour of small groups of fibroblasts

1965 ◽  
Vol 162 (988) ◽  
pp. 289-302 ◽  
Keyword(s):  
Small Groups ◽  
Control Groups ◽  
Isolated Cells ◽  
Daughter Cells ◽  
A Cell ◽  
Locomotory Behaviour ◽  
Chick Embryo Heart ◽  
Embryo Heart ◽  
Random Nature ◽  
Chemotactic Gradient

Small groups of two to four fibroblasts at the periphery of outgrowths from cultured explants of chick embryo heart were isolated from their neighbours by sweeping away the nearby cells. The groups and the explants were left attached to the glass substrate, undisturbed. The behaviour of the isolated cells was photographically recorded during about 8 h of further culture. The cells of these groups dispersed, though not as a rule so far as to lose all mutual contacts, the dispersal being counterbalanced by the addition of new cells through mitosis. The accompanying changes in speed of locomotion, and the non-random nature of the spreading, are interpreted in terms of the effects of contacts between the cells. During the first four hours after isolation, but not thereafter, the cells of the groups on the average moved slowly away from the explant. Control groups in an intact outgrowth moved away faster and with no diminution of speed during the period of observation. The movement of the isolated groups can be partly accounted for by the tendency of cells to conserve for a time the direction of their movement before isolation; and by a strong reluctance of the isolated cells to move across the area, from which cells had been scraped away, that lay between the group and the explant. A new outgrowth of the residual sheet of cells still connected to the explant, however, advanced across this area, approaching and in most cases overhauling the isolated group. It is concluded that a chemotactic gradient around the explant is unlikely to play any significant part in the outward movement of fibroblasts from an explant in tissue culture. The cells of the isolated groups underwent an outburst of mitosis about 3 h after isolation. Mitoses in these relatively free cells are oriented in relation to the polarity of the cell before division. Locomotion of the daughter-cells tends to be faster than usual for at least 2 h after a cell divides.

scholarly journals e-Book on plant virus infection—a cell biology perspective

2013 ◽  
Vol 4 ◽  
Author(s):  
Jean-François Laliberté ◽  
Peter Moffett ◽  
Hélène Sanfaçon ◽  
Aiming Wang ◽  
Richard S. Nelson ◽  
...  
Keyword(s):  
Virus Infection ◽  
Cell Biology ◽  
Plant Virus ◽  
A Cell

(A) Cell biology of epithelia

1991 ◽  
Vol 7 (3) ◽  
pp. 313-338 ◽  
Author(s):  
Martin Mackay ◽  
Ian Williamson ◽  
John Hastewell
Keyword(s):  
Cell Biology ◽  
A Cell

A highly repetitive DNA family is dispersed in small clusters throughout the genome

1986 ◽  
Vol 10 (6) ◽  
pp. 486 ◽  
Author(s):  
R BATISTONI ◽  
R VIGNALI ◽  
A NEGRONI ◽  
F CREMISI ◽  
G BARSACCHIPILONE
Keyword(s):  
Repetitive Dna ◽  
Highly Repetitive Dna ◽  
Small Clusters

Autonomously replicating episomes contain mdr1 genes in a multidrug-resistant human cell line

1989 ◽  
Vol 9 (1) ◽  
pp. 109-115
Author(s):  
J C Ruiz ◽  
K H Choi ◽  
D D von Hoff ◽  
I B Roninson ◽  
G M Wahl
Keyword(s):  
Gamma Irradiation ◽  
Gene Amplification ◽  
Human Tumor ◽  
Human Cell Line ◽  
Multidrug Resistant ◽  
Cross Resistance ◽  
Chromosomal Dna ◽  
A Cell ◽  
Shift Experiment ◽  
Voltage Gradients

Gene amplification in human tumor cells is frequently mediated by extrachromosomal elements (e.g., double minute chromosomes [DMs]). Recent experiments have shown that DMs can be formed from smaller, submicroscopic circular precursors referred to as episomes (S. M. Carroll, M. L. DeRose, P. Gaudray, C. M. Moore, D. R. Needham-Vandevanter, D. D. Von Hoff and G. M. Wahl, Mol. Biol. 8:1525-1533, 1988). To investigate whether episomes are generally involved as intermediates in gene amplification, we determined whether they mediate the amplification of the mdr1 gene, which when overexpressed engenders cross resistance to multiple lipophilic drugs. A variety of methods including electrophoresis of undigested DNAs in high-voltage gradients, NotI digestion, and production of double-strand breaks by gamma irradiation were used to distinguish between mdr1 sequences amplified on submicroscopic circular molecules and those amplified within DMs or chromosomal DNA. The gamma-irradiation procedure provides a new method for detecting and determining the size of circular molecules from 50 kilobases (kb) to greater than 1,000 kb. These methods revealed that some of the amplified mdr1 genes in vinblastine-resistant KB-V1 cells are contained in supercoiled circular molecules of approximately 600 and approximately 750 kb. Analysis of the replication of these molecules by a Meselson-Stahl density shift experiment demonstrated that they replicate approximately once in a cell cycle. The data lend further support to a model for gene amplification in which DMs are generally formed from smaller, autonomously replicating precursors.

scholarly journals Ser68 phosphoregulation is essential for CENP-A deposition, centromere function and viability in mice

2021 ◽  
Author(s):  
Yuting Liu ◽  
Kehui Wang ◽  
Li Huang ◽  
Jicheng Zhao ◽  
Xinpeng Chen ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Viability ◽  
Histone H3 ◽  
Dependent Manner ◽  
Centromere Function ◽  
Histone H3 Variant ◽  
A Cell ◽  
Cycle Dependent ◽  
Spatiotemporal Control

Centromere identity is defined by nucleosomes containing CENP-A, a histone H3 variant. The deposition of CENP-A at centromeres is tightly regulated in a cell-cycle-dependent manner. We previously reported that the spatiotemporal control of centromeric CENP-A incorporation is mediated by the phosphorylation of CENP-A Ser68. However, a recent report argued that Ser68 phosphoregulation is dispensable for accurate CENP-A loading. Here, we report that the substitution of Ser68 of endogenous CENP-A with either Gln68 or Glu68 severely impairs CENP-A deposition and cell viability. We also find that mice harboring the corresponding mutations are lethal. Together, these results indicate that the dynamic phosphorylation of Ser68 ensures cell-cycle-dependent CENP-A deposition and cell viability.

scholarly journals Auxin/AID versus conventional knockouts: distinguishing the roles of CENP-T/W in mitotic kinetochore assembly and stability

Open Biology ◽  
2016 ◽  
Vol 6 (1) ◽  
pp. 150230 ◽  
Author(s):  
Laura Wood ◽  
Daniel G. Booth ◽  
Giulia Vargiu ◽  
Shinya Ohta ◽  
Flavia deLima Alves ◽  
...  
Keyword(s):  
Protein Function ◽  
Mitotic Chromosomes ◽  
Protein Levels ◽  
Kinetochore Assembly ◽  
Rnai Technology ◽  
Gradual Loss ◽  
A Cell ◽  
Stable Association ◽  
Or Gene ◽  
Regulated Gene Expression

Most studies using knockout technologies to examine protein function have relied either on shutting off transcription (conventional conditional knockouts with tetracycline-regulated gene expression or gene disruption) or destroying the mature mRNA (RNAi technology). In both cases, the target protein is lost at a rate determined by its intrinsic half-life. Thus, protein levels typically fall over at least 1–3 days, and cells continue to cycle while exposed to a decreasing concentration of the protein. Here we characterise the kinetochore proteome of mitotic chromosomes isolated from a cell line in which the essential kinetochore protein CENP-T is present as an auxin-inducible degron (AID) fusion protein that is fully functional and able to support the viability of the cells. Stripping of the protein from chromosomes in early mitosis via targeted proteasomal degradation reveals the dependency of other proteins on CENP-T for their maintenance in kinetochores. We compare these results with the kinetochore proteome of conventional CENP-T/W knockouts. As the cell cycle is mostly formed from G1, S and G2 phases a gradual loss of CENP-T/W levels is more likely to reflect dependencies associated with kinetochore assembly pre-mitosis and upon entry into mitosis. Interestingly, a putative super-complex involving Rod-Zw10-zwilch (RZZ complex), Spindly, Mad1/Mad2 and CENP-E requires the function of CENP-T/W during kinetochore assembly for its stable association with the outer kinetochore, but once assembled remains associated with chromosomes after stripping of CENP-T during mitosis. This study highlights the different roles core kinetochore components may play in the assembly of kinetochores (upon entry into mitosis) versus the maintenance of specific components (during mitosis).

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