scholarly journals Efficient combination of large DNA in vitro: in gel site specific recombination (IGSSR) of PAC fragments containing alpha satellite DNA and the human HPRT gene locus

1997 ◽  
Vol 25 (11) ◽  
pp. 2241-2243 ◽  
Author(s):  
D Schindelhauer
Keyword(s):  
Satellite Dna ◽  
Gene Locus ◽  
Alpha Satellite ◽  
Hprt Gene ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Alpha Satellite Dna ◽  
Human Hprt Gene

scholarly journals Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere.

1992 ◽  
Vol 116 (5) ◽  
pp. 1081-1093 ◽  
Author(s):  
A F Pluta ◽  
N Saitoh ◽  
I Goldberg ◽  
W C Earnshaw
Keyword(s):  
Satellite Dna ◽  
Centromeric Heterochromatin ◽  
Amino Terminus ◽  
Major Protein ◽  
Alpha Satellite ◽  
Alpha Satellite Dna ◽  
Human Centromere ◽  
Bacterial Enzyme

We have combined in vivo and in vitro approaches to investigate the function of CENP-B, a major protein of human centromeric heterochromatin. Expression of epitope-tagged deletion derivatives of CENP-B in HeLa cells revealed that a single domain less than 158 residues from the amino terminus of the protein is sufficient to localize CENP-B to centromeres. Centromere localization was abolished if as few as 28 amino acids were removed from the amino terminus of CENP-B. The centromere localization signal of CENP-B can function in an autonomous fashion, relocating a fused bacterial enzyme to centromeres. The centromere localization domain of CENP-B specifically binds in vitro to a subset of alpha-satellite DNA monomers. These results suggest that the primary mechanism for localization of CENP-B to centromeres involves the recognition of a DNA sequence found at centromeres. Analysis of the distribution of this sequence in alpha-satellite DNA suggests that CENP-B binding may have profound effects on chromatin structure at centromeres.

scholarly journals CENP-C binds the alpha-satellite DNA in vivo at specific centromere domains

2002 ◽  
Vol 115 (11) ◽  
pp. 2317-2327 ◽  
Author(s):  
Valeria Politi ◽  
Giovanni Perini ◽  
Stefania Trazzi ◽  
Artem Pliss ◽  
Ivan Raska ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Satellite Dna ◽  
Alpha Satellite ◽  
Alpha Satellite Dna ◽  
Human Centromere ◽  
Ultrastructural Characterization ◽  
First Time

CENP-C is a fundamental component of the centromere, highly conserved among species and necessary for the proper assembly of the kinetochore structure and for the metaphase-anaphase transition. Although CENP-C can bind DNA in vitro,the identification of the DNA sequences associated with it in vivo and the significance of such an interaction have been, until now, elusive. To address this problem we took advantage of a chromatin-immunoprecipitation procedure and applied this technique to human HeLa cells. Through this approach we could establish that: (1) CENP-C binds the alpha-satellite DNA selectively; (2) the CENP-C region between amino acids 410 and 537, previously supposed to contain a DNA-binding domain, is indeed required to perform such a function in vivo;and (3) the profile of the alpha-satellite DNA associated with CENP-C is essentially identical to that recognized by CENP-B. However, further biochemical and ultrastructural characterization of CENP-B/DNA and CENP-C/DNA complexes, relative to their DNA components and specific spatial distribution in interphase nuclei, surprisingly reveals that CENP-C and CENP-B associate with the same types of alpha-satellite arrays but in distinct non-overlapping centromere domains. Our results, besides extending previous observations on the role of CENP-C in the formation of active centromeres, show, for the first time, that CENP-C can associate with the centromeric DNA sequences in vivo and, together with CENP-B, defines a highly structured organization of the alpha-satellite DNA within the human centromere.

On the mode of evolution of alpha satellite DNA in human populations

1991 ◽  
Vol 33 (1) ◽  
pp. 42-48 ◽  
Author(s):  
B. Marçais ◽  
J. P. Charlieu ◽  
B. Allain ◽  
E. Brun ◽  
M. Bellis ◽  
...  
Keyword(s):  
Satellite Dna ◽  
Human Populations ◽  
Alpha Satellite ◽  
Alpha Satellite Dna

Site-Specific Recombination Promoted in Vitro by the FLP Protein of the Yeast Two-Micron Plasmid

1986 ◽  
pp. 397-405
Author(s):  
Julie F. Senecoff ◽  
Robert C. Bruckner ◽  
Leslie Meyer-Leon ◽  
Cynthia A. Gates ◽  
Elizabeth Wood ◽  
...  
Keyword(s):  
Site Specific ◽  
Site Specific Recombination

scholarly journals Characterization of the Chromosome Dimer Resolution Site in Caulobacter crescentus

2019 ◽  
Vol 201 (24) ◽  
Author(s):  
Ali Farrokhi ◽  
Hua Liu ◽  
George Szatmari
Keyword(s):  
Cell Division ◽  
Caulobacter Crescentus ◽  
Consensus Sequence ◽  
Markov Modeling ◽  
Control Of Gene Expression ◽  
Site Specific ◽  
Content Type ◽  
Site Specific Recombination ◽  
Resolution System

ABSTRACT Chromosome dimers occur in bacterial cells as a result of the recombinational repair of DNA. In most bacteria, chromosome dimers are resolved by XerCD site-specific recombination at the dif (deletion-induced filamentation) site located in the terminus region of the chromosome. Caulobacter crescentus, a Gram-negative oligotrophic bacterium, also possesses Xer recombinases, called CcXerC and CcXerD, which have been shown to interact with the Escherichia coli dif site in vitro. Previous studies on Caulobacter have suggested the presence of a dif site (referred to in this paper as dif1CC), but no in vitro data have shown any association with this site and the CcXer proteins. Using recursive hidden Markov modeling, another group has proposed a second dif site, which we call dif2CC, which shows more similarity to the dif consensus sequence. Here, by using a combination of in vitro experiments, we compare the affinities and the cleavage abilities of CcXerCD recombinases for both dif sites. Our results show that dif2CC displays a higher affinity for CcXerC and CcXerD and is bound cooperatively by these proteins, which is not the case for the original dif1CC site. Furthermore, dif2CC nicked substrates are more efficiently cleaved by CcXerCD, and deletion of the site results in about 5 to 10% of cells showing an altered cellular morphology. IMPORTANCE Bacteria utilize site-specific recombination for a variety of purposes, including the control of gene expression, acquisition of genetic elements, and the resolution of dimeric chromosomes. Failure to resolve dimeric chromosomes can lead to cell division defects in a percentage of the cell population. The work presented here shows the existence of a chromosomal resolution system in C. crescentus. Defects in this resolution system result in the formation of chains of cells. Further understanding of how these cells remain linked together will help in the understanding of how chromosome segregation and cell division are linked in C. crescentus.

scholarly journals Xer-mediated site-specific recombination in vitro.

1996 ◽  
Vol 15 (5) ◽  
pp. 1172-1181 ◽  
Author(s):  
S. D. Colloms ◽  
R. McCulloch ◽  
K. Grant ◽  
L. Neilson ◽  
D. J. Sherratt
Keyword(s):  
Site Specific ◽  
Site Specific Recombination
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