Adenylation and ADP-ribosylation in the mouse 1-cell embryo

Development ◽  
1979 ◽  
Vol 49 (1) ◽  
pp. 139-152
Author(s):  
R. J. Young ◽  
K. Sweeney
Keyword(s):  
Dna Synthesis ◽  
Snake Venom ◽  
Trichloroacetic Acid ◽  
Actinomycin D ◽  
Adp Ribosylation ◽  
Snake Venom Phosphodiesterase ◽  
Insoluble Material ◽  
Cell Embryo

The incorporation of [3H]adenosine into cold trichloroacetic acid (TCA) insoluble material by the mouse 1-cell embryo has been studied. Incorporation of label was high immediately after fertilization, then decreased over the next 7 h with the sharpest decline occurring 3—5 h after fertilization. A small maximum was observed at the time of pronuclear DNA synthesis. Actinomycin D at a concentration which inhibited the cleavage of 1-cell embryos by 50 % had little effect on this incorporation, which in the period 1—6 h post-fertilization was shown by autoradiography to be confined to the ooplasm of the newly fertilized ovum. [3H]Adenosine and poly ([H]A) were released from embryo RNA labelled 1—3 h after fertilization with [3H]adenosine by digestion with a mixture of ribonucleases A and T1. The poly ([3H]A) segments were hydrolysed by alkali to 3′-[3H]AMP and [3H]adenosine ([3H]AMP/[3H]adenosine = 5/1), and by snake venom phosphodiesterase to 5′-[3H]AMP but very little [3H]adenosine. These results suggest that adenylation of RNA occurs soon after fertilization, that this is a cytoplasmic event, and that most of the newly synthesized poly ([3H]A) segments are joined to pre-existing poly (A) tracts. The unusual polynucleotide, poly (ADP-ribose), identified by its resistance to alkali and the release of 2′-(5″-phosphoribosyl)-5′[3H]AMP on incubation with snake venom phosphodiesterase, was also found in the ribonuclease digest.

The Synthesis of Nucleic Acids in Bacillus subtilis Infected with Phage PBS 1

1973 ◽  
Vol 51 (9) ◽  
pp. 1219-1224 ◽  
Author(s):  
B. K. Rima ◽  
I. Takahashi
Keyword(s):  
Bacillus Subtilis ◽  
Nucleic Acids ◽  
Dna Synthesis ◽  
Trichloroacetic Acid ◽  
Actinomycin D ◽  
The Other ◽  
Phage Dna ◽  
Infected Cells ◽  
Phage Growth ◽  
A New Species

Unlike other phage systems, the development of PBS 1 was found to be insensitive to rifamycin SV. The incorporation of 3H-uridine into trichloroacetic acid precipitable and alkali-labile material (RNA), in PBS-1-infected cells, was greatly reduced by rifamycin. Observations that RNA synthesized in the presence of rifamycin was hybridizable exclusively with the phage DNA and that actinomycin D inhibited the phage growth indicated that the synthesis of a new species of RNA was required for the development of PBS 1. The host DNA synthesis was reduced to a very low level 5 min after infection. The phage DNA synthesis was also apparently reduced markedly by rifamycin when determined with 3H-uridine as labelling material. On the other hand, rifamycin did not affect the incorporation of 3H-deoxycytidine into the phage DNA, suggesting that phage DNA synthesis was in fact insensitive to rifamycin. It is not clear how rifamycin inhibits the incorporation of 3H-uridine into nucleic acids in PBS-1-infected cells.

scholarly journals Stereochemistry of hydrolysis by snake venom phosphodiesterase.

1979 ◽  
Vol 254 (16) ◽  
pp. 7476-7478 ◽  
Author(s):  
P.M. Burgers ◽  
F. Eckstein ◽  
D.H. Hunneman
Keyword(s):  
Snake Venom ◽  
Snake Venom Phosphodiesterase

scholarly journals Arginine deprivation in KB cells: I. Effect on cell cycle progress

1977 ◽  
Vol 75 (3) ◽  
pp. 881-888 ◽  
Author(s):  
AS Weissfeld ◽  
H Rouse
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Tritiated Thymidine ◽  
Experimental Period ◽  
Cell Multiplication ◽  
Starvation Period ◽  
Kb Cells ◽  
Insoluble Material ◽  
Cell Cycle Inhibition ◽  
Cellular Dna

When exponentially growing KB cells were deprived of arginine, cell multiplication ceased after 12 h but viability was maintained throughout the experimental period (42-48 h). Although tritiated thymidine ([(3)H]TdR) incorporation into acid-insoluble material declined to 5 percent of the initial rate, the fraction of cells engaged in DNA synthesis, determined by autoradiography, remained constant throughout the starvation period and approximately equal to the synthesizing fraction in exponentially growing controls (40 percent). Continous [(3)H]TdR-labeling indicated that 80 percent of the arginine-starved cells incorporated (3)H at some time during a 48-h deprivation period. Thus, some cells ceased DNA synthesis, whereas some initially nonsynthesizing cells initiated DNA synthesis during starvation. Flow microfluorometric profiles of distribution of cellular DNA contents at the end of the starvation period indicated that essentially no cells had a 4c or G2 complement. If arginine was restored after 30 h of starvation, cultures resumed active, largely asynchronous division after a 16-h lag. Autoradiographs of metaphase figures from cultures continuously labeled with [(3)H]TdR after restoration indicated that all cells in the culture underwent DNA synthesis before dividing. It was concluded that the majority of cells in arginine-starved cultures are arrested in neither a normal G1 nor G2. It is proposed that for an exponential culture, i.e. from most positions in the cell cycle, inhibition of cell growth after arginine with withdrawal centers on the ability of cells to complete replication of their DNA.

scholarly journals Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

1983 ◽  
Vol 3 (1) ◽  
pp. 70-81 ◽  
Author(s):  
C D Scher ◽  
R L Dick ◽  
A P Whipple ◽  
K L Locatell
Keyword(s):  
Growth Factor ◽  
Dna Synthesis ◽  
Cell Lines ◽  
Actinomycin D ◽  
Platelet Derived Growth Factor ◽  
Cell Protein ◽  
Transformed Cells ◽  
Antigenic Determinants ◽  
3T3 Cells ◽  
Constitutive Synthesis

The platelet-derived growth factor (PDGF) stimulates density-arrested BALB/c-3T3 cells to synthesize a protein (pII; Mr, 35,000) that is constitutively synthesized by spontaneously transformed BALB/c-3T3 (ST2-3T3) cells which do not require PDGF for growth. Antisera against a major excreted protein family (MEP) of retrovirus-transformed cells quantitatively precipitated cellular pII. PDGF-stimulated pII has the same molecular weight, a similar charge, and similar antigenic determinants as authentic MEP isolated from ST2-3T3 or retrovirus-transformed cells. MEP represented about 2% of the nonnuclear proteins synthesized by ST2-3T3 cells and 0.3 to 0.6% of the proteins synthesized by PDGF-treated BALB/c-3T3 cells, a three- to sixfold increase over the background. In BALB/c-3T3 cells, less PDGF was required for pII (MEP) synthesis than for DNA synthesis. PDGF induced a selective increase in pII (MEP) within 40 min. Such preferential synthesis was inhibited by brief treatment with actinomycin D, suggesting a requirement for newly formed RNA. The constitutive synthesis of pII (MEP) by ST2-3T3 cells was not inhibited by actinomycin D. Five spontaneously or chemical carcinogen-transformed tumorigenic BALB/c-3T3 cell lines were studied; they neither required PDGF for growth nor responded to it. These cell lines became arrested at confluence with a G1 DNA content. Each of these independently isolated lines synthesized pII (MEP) constitutively. Thus, the synthesis of pII (MEP) may be required, but is not sufficient, for PDGF-modulated DNA synthesis.

Synthesis of phosphoramidate analogs of ribodinucleoside phosphates

1979 ◽  
Vol 44 (3) ◽  
pp. 792-798 ◽  
Author(s):  
Alexei V. Azhayev ◽  
Alexander A. Krayevsky ◽  
Jiří Smrt
Keyword(s):  
Snake Venom ◽  
Pancreatic Ribonuclease ◽  
Carbon Tetrabromide ◽  
Snake Venom Phosphodiesterase

Cytidine 5'-phosphate (Ia) reacts with 1,1'-carbonyldiimidazole under the formation of 2',3'-O-carbonylcytidine 5'-phosphorimidazolidate (IIa) which affords (by the action of 3'-amino-3'-deoxyadenosine (IIIa)) 3'-deoxyadenosine-3'-amidophosphoryl-(3' → 5')-cytidine (IVa) and (by the action of 3'-amino-3-deoxy-N6-dimethyladenosine (IIIc)) 3'-deoxy-N6-dimethyladenosine-3'-amidophosphoryl-(3' → 5')-cytidine (IVc). Starting from adenosine 5'-phosphate (Ib) and 3'-amino-3'-deoxycytidine (IIIb) or the substance IIIc, 3'-deoxycytidine-3'-amidophosphoryl-(3' → 5')-adenosine (IVb) or 3'-deoxy-N6-dimethyladenosine-3'-amidophosphoryl-(3' → 5')-adenosine (IVd) were prepared. 6-Azauridine (V) was transformed, by the action of triphenylphosphine, lithium azide and carbon tetrabromide, followed by the action of triphenylphosphine and ammonia, to 5'-amino-5'-deoxy-6-azauridine (VI). The substance VI was transformed by the action of 2',3'-O-carbonyl-adenosine 5'-phosphorimidazolidate (IIb), to adenylyl-(5' → 5')-5'-amino-5'-deoxy-6-azauridine (VII). The compound IVb is not degraded by snake venom and spleen phosphodiesterases and is degraded by pancreatic ribonuclease to adenosine and the compound IIIb. The compound VII is degraded by snake venom phosphodiesterase to adenosine and the compound VI.

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