scholarly journals Mapping of replication initiation sites in human ribosomal DNA by nascent-strand abundance analysis.

1995 ◽  
Vol 15 (5) ◽  
pp. 2482-2489 ◽  
Author(s):  
Y Yoon ◽  
J A Sanchez ◽  
C Brun ◽  
J A Huberman
Keyword(s):  
Ribosomal Dna ◽  
Gel Electrophoresis ◽  
Repeat Unit ◽  
Low Frequency ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Replication Initiation ◽  
Size Analysis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

New techniques for mapping mammalian DNA replication origins are needed. We have modified the existing nascent-strand size analysis technique (L. Vassilev and E.M. Johnson, Nucleic Acids Res. 17:7693-7705, 1989) to provide an independent means of studying replication initiation sites. We call the new method nascent-strand abundance analysis. We confirmed the validity of this method with replicating simian virus 40 DNA as a model. We then applied nascent-strand abundance and nascent-strand size analyses to mapping of initiation sites in human (HeLa) ribosomal DNA (rDNA), a region previously examined exclusively by two-dimensional gel electrophoresis methods (R.D. Little, T.H.K. Platt, and C.L. Schildkraut, Mol. Cell. Biol. 13:6600-6613, 1993). Our results partly confirm those obtained by two-dimensional gel electrophoresis techniques. Both studies suggest that replication initiates at relatively high frequency a few kilobase pairs upstream of the transcribed region and that many additional low-frequency initiation sites are distributed through most of the remainder of the ribosomal DNA repeat unit.

scholarly journals Simian virus 40 and polyoma virus induce synthesis of heat shock proteins in permissive cells.

1983 ◽  
Vol 3 (1) ◽  
pp. 1-8 ◽  
Author(s):  
E W Khandjian ◽  
H Türler
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Gel Electrophoresis ◽  
Mammalian Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Polyoma Virus ◽  
Two Dimensional Gel Electrophoresis ◽  
Mouse Cells ◽  
Stimulation Of

During the lytic infection of monkey and mouse cells with simian virus 40 and polyoma virus, respectively, the preferentially increased synthesis of two host proteins of 92,000 and 72,000 Mr was observed by 15 to 20 h after infection besides the general stimulation of most cellular proteins. The incubation of uninfected monkey and mouse cell cultures for 30 to 60 min at 43.5 degrees C induced the enhanced synthesis of at least three proteins of 92,000, 72,000 and 70,000 Mr, the last one being the major heat shock protein of mammalian cells. Two-dimensional gel electrophoresis and partial proteolytic digestion confirmed that the same 92,000- and 72,000-Mr proteins are stimulated by virus infection and thermal treatment. In simian virus 40-infected CV-1 cells, we also observed the weak stimulation of a 70,000-Mr protein comigrating in gel electrophoresis with the major heat shock protein. The 92,000-, 72,000- and 70,000-Mr proteins of monkey cells are structurally very similar to the corresponding proteins of mouse cells. In immunoprecipitations, no specific association of these proteins to simian virus 40 T antigens was noticed.

Two-dimensional gel electrophoresis of human brain proteins. III. Genetic and non-genetic variations in 145 brains.

1982 ◽  
Vol 28 (4) ◽  
pp. 798-804 ◽  
Author(s):  
D E Comings
Keyword(s):  
Human Brain ◽  
Gel Electrophoresis ◽  
Low Frequency ◽  
Genetic Variations ◽  
Genetic Mutations ◽  
Two Dimensional ◽  
Chi Square ◽  
Two Dimensional Gel Electrophoresis ◽  
Brain Proteins ◽  
Chi Square Analysis

Abstract To determine the frequency of genetic mutations, polymorphisms, and non-genetic variation in the major human brain proteins, I examined, by equilibrium two-dimensional gel electrophoresis, 145 brains from patients dying of a wide variety of psychiatric, neurological, and non-neurological disorders. Of 176 polypeptides screened, there was one polymorphism of glial fibrillary acidic protein (GFAP-Duarte). Chi square analysis indicated it was non-randomly distributed among different diseases. A possible mutation associated with Joseph's disease is being further investigated. Three examples of a possible mutation of protein 8c:1 were noted. No other genetic mutations were observed. This low frequency of polymorphisms is consistent with results for two-dimensional gel analysis of other tissue and species. The numerous non-genetic variations are described.

Simian virus 40 and polyoma virus induce synthesis of heat shock proteins in permissive cells

1983 ◽  
Vol 3 (1) ◽  
pp. 1-8
Author(s):  
E W Khandjian ◽  
H Türler
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Gel Electrophoresis ◽  
Mammalian Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Polyoma Virus ◽  
Two Dimensional Gel Electrophoresis ◽  
Mouse Cells ◽  
Stimulation Of

During the lytic infection of monkey and mouse cells with simian virus 40 and polyoma virus, respectively, the preferentially increased synthesis of two host proteins of 92,000 and 72,000 Mr was observed by 15 to 20 h after infection besides the general stimulation of most cellular proteins. The incubation of uninfected monkey and mouse cell cultures for 30 to 60 min at 43.5 degrees C induced the enhanced synthesis of at least three proteins of 92,000, 72,000 and 70,000 Mr, the last one being the major heat shock protein of mammalian cells. Two-dimensional gel electrophoresis and partial proteolytic digestion confirmed that the same 92,000- and 72,000-Mr proteins are stimulated by virus infection and thermal treatment. In simian virus 40-infected CV-1 cells, we also observed the weak stimulation of a 70,000-Mr protein comigrating in gel electrophoresis with the major heat shock protein. The 92,000-, 72,000- and 70,000-Mr proteins of monkey cells are structurally very similar to the corresponding proteins of mouse cells. In immunoprecipitations, no specific association of these proteins to simian virus 40 T antigens was noticed.

Two-dimensional gel electrophoretic method for mapping DNA replicons

1988 ◽  
Vol 8 (4) ◽  
pp. 1408-1413 ◽  
Author(s):  
K A Nawotka ◽  
J A Huberman
Keyword(s):  
Gel Electrophoresis ◽  
Replication Origin ◽  
Replication Fork ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Electrophoretic Method ◽  
Short Probe

We describe in detail a method which allows determination of the directions of replication fork movement through segments of DNA for which cloned probes are available. The method uses two-dimensional neutral-alkaline agarose gel electrophoresis followed by hybridization with short probe sequences. The nascent strands of replicating molecules form an arc separated from parental and nonreplicating strands. The closer a probe is to its replication origin or to the origin-proximal end of its restriction fragment, the shorter the nascent strands that are detected by the probe. The use of multiple probes allows determination of directions of replication fork movement, as well as locations of origins and termini. In this study, we used simian virus 40 as a model to demonstrate the feasibility of the method, and we discuss its applicability to other systems.

scholarly journals Two-dimensional gel electrophoretic method for mapping DNA replicons.

1988 ◽  
Vol 8 (4) ◽  
pp. 1408-1413 ◽  
Author(s):  
K A Nawotka ◽  
J A Huberman
Keyword(s):  
Gel Electrophoresis ◽  
Replication Origin ◽  
Replication Fork ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Electrophoretic Method ◽  
Short Probe

We describe in detail a method which allows determination of the directions of replication fork movement through segments of DNA for which cloned probes are available. The method uses two-dimensional neutral-alkaline agarose gel electrophoresis followed by hybridization with short probe sequences. The nascent strands of replicating molecules form an arc separated from parental and nonreplicating strands. The closer a probe is to its replication origin or to the origin-proximal end of its restriction fragment, the shorter the nascent strands that are detected by the probe. The use of multiple probes allows determination of directions of replication fork movement, as well as locations of origins and termini. In this study, we used simian virus 40 as a model to demonstrate the feasibility of the method, and we discuss its applicability to other systems.

Diversity of Glycoprotein Deficiencies in Glanzmann’s Thrombasthenia

1985 ◽  
Vol 54 (03) ◽  
pp. 626-629 ◽  
Author(s):  
M Meyer ◽  
F H Herrmann
Keyword(s):  
High Resolution ◽  
Gel Electrophoresis ◽  
Type Ii ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Structural Variant ◽  
Glanzmann’S Thrombasthenia ◽  
Crossed Immunoelectrophoresis ◽  
Glanzmann's Thrombasthenia ◽  
Platelet Proteins

SummaryThe platelet proteins of 9 thrombasthenic patients from 7 families were analysed by high resolution two-dimensional gel electrophoresis (HR-2DE) and crossed immunoelectrophoresis (CIE). In 7 patients both glycoproteins (GPs) IIb and Ilia were absent or reduced to roughly the same extent. In two related patients only a trace of GP Ilb-IIIa complex was detected in CIE, but HR-2DE revealed a glycopeptide in the position of GP Ilia in an amount comparable to type II thrombasthenia. This GP Ilia-like component was neither recognized normally by anti-GP Ilb-IIIa antibodies nor labeled by surface iodination. In unreduced-reduced two-dimensional gel electrophoresis two components were observed in the region of GP Ilia. The assumption of a structural variant of GP Ilia in the two related patients is discussed.

Comparative Two-Dimensional Gel Electrophoresis of Trypanosoma cruzi Mammalian-Stage Forms in an Alkaline pH Range

2015 ◽  
Vol 22 (12) ◽  
pp. 1066-1075 ◽  
Author(s):  
Adriana Magalhães ◽  
Rayner Queiroz ◽  
Izabela Bastos ◽  
Jaime Santana ◽  
Marcelo Sousa ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Alkaline Ph ◽  
Two Dimensional Gel Electrophoresis ◽  
Ph Range
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