Isolation of a distinct pool of polyribosomes from nuclei of adenovirus-infected HeLa cells

Chatterjee, Nando K.; Dickerman, Herbert W.; Beach, Thaisa A.

doi:10.1016/0003-9861(77)90436-2

Cited by 16 publications

(6 citation statements)

References 21 publications

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“…In a similar study we (Chatterjee et al, 1977; N. K. Chatterjee, C. Tuchowski, G. E. Eagan & T. M. Haley, unpublished work) found a distinct population of polyribosomes containing functional mRNA in adenovirus-infected HeLa-cell nuclei. These polyribosomes were released from detergent-washed intact nuclei by poly(U) or other polynucleotides.…”

supporting

confidence: 66%

“…Procedures have been described for the growth of HeLa cells (S3) in suspension cultures, the growth of type-2 adenovirus in these cells, and purification of the virus (Maizel et al, 1968), as well as for infecting HeLa cells with 3000-4000 adenovirus particles/cell and preparing detergent (Triton N101)-washed nuclei and the postnuclear cytoplasmic supernatant (Chatterjee et al, 1977). Labelling, isolation and purification of nuclear and cytoplasmic polyribosomes HeLa cells were grown in a phosphate-free Joklik-modified F-13 medium (Grand Island Bio-logical) overnight, infected with the virus, and grown in the same medium.…”

Section: Virus-infected Cells: Preparation Of Nuclei and Postnuclear mentioning

confidence: 99%

“…The procedure of Goidl et al (1975) was modified (Chatterjee et al, 1977) to release nuclear polyribosomes from the detergent-washed nuclei. Briefly, nuclei (equivalent to 15-20mg of protein) were incubated in 6ml of 10mM-Tris/HCl (pH7.5)/125mM-sucrose containing lmg of poly(U) for 10min at 0°C.…”

Section: Virus-infected Cells: Preparation Of Nuclei and Postnuclear mentioning

confidence: 99%

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Differences in sequence content of nuclear and cytoplasmic polyribosomal RNA from adenovirus-infected cells

Chatterjee¹,

Tuchowski²,

Eagan³

et al. 1984

Biochemical Journal

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RNA molecules from nuclear and cytoplasmic polyribosomes of adenovirus-infected HeLa cells were compared by hybridization to analyse the sequence content. Nuclear polyribosomes were released by exposure of intact detergent-washed nuclei to poly(U) and purified. Cytoplasmic polyribosomes were also purified from the same cells. To show that nuclear polyribosomes contain ribosomes linked by mRNA, polyribosomes were labelled with methionine and uridine in the presence of actinomycin D in adenovirus-infected cells. Purified nuclear polyribosomes were treated with EDTA under conditions which dissociate polyribosomes into ribosomes and subunits with a simultaneous release of mRNA, and sedimented. The treatment dissociated these polyribosomes, releasing the mRNA from them. Radiolabelled total RNA from each polyribosome population was fractionated in sucrose gradients into several pools or hybridized to intact adenovirus DNA to select virus-specific RNA. Sucrose-gradient-fractionated pool-3 RNA (about 28S) and virus-specific RNA were then hybridized to fragments of adenovirus DNA cleaved by restriction endonucleases SmaI, HindIII and EcoRI by the Southern-blot technique and by filter hybridization. The results showed that nuclear RNA contained sequences, from about 0 to 18 map units, which were essentially absent from cytoplasmic RNA. Furthermore, the amount of virus-specific RNA for a particular sequence was also different in the two populations.

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supporting

confidence: 66%

Section: Virus-infected Cells: Preparation Of Nuclei and Postnuclear mentioning

confidence: 99%

Section: Virus-infected Cells: Preparation Of Nuclei and Postnuclear mentioning

confidence: 99%

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Differences in sequence content of nuclear and cytoplasmic polyribosomal RNA from adenovirus-infected cells

Chatterjee¹,

Tuchowski²,

Eagan³

et al. 1984

Biochemical Journal

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“…It was also not due to the entry of some essential factor through the pore as thapsigargin, a drug that inhibits passive diffusion of molecules as small as 10 kDa through the pore (Greber and Gerace 1995), had no effect on the nuclear signal. And even if cytoplasmic ribosomes did enter to initiate on nuclear message, they could not be responsible for the signal as aurintricarboxylic acid, which inhibits initiation and not elongation (Chatterjee et al 1977), had no effect.…”

Section: Nuclear Translationmentioning

confidence: 98%

The path that RNA takes from the nucleus to the cytoplasm: a trip with some surprises

Iborra

2002

Histochem Cell Biol

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“…During our investigation on ribosomes of adenovirus-infected HeLa cells, we observed that ribosomes of nuclear and cytoplasmic origin containing translatable mRNA have significant cyclic nucleotide-independent protein kinase activity and a variety of particle-associated endogenous acceptor proteins (Chatterjee et al, 1977(Chatterjee et al, , 1979. Initial experiments suggested that the kinases and the acceptor proteins may be different in the two populations.…”

mentioning

confidence: 99%

Cyclic nucleotide-independent protein kinases bound to cytoplasmic and nuclear polyribosomes in non-infected and adenovirus-infected HeLa cells

Sarma

Chatterjee

1980

Biochemical Journal

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Cyclic nucleotide-independent protein kinase activity bound to cytoplasmic and nuclear polyribsomes from non-infected and adenovirus-infected HeLa cells was compared. The enzymes catalysed the incorporation of phosphate from gamma-(32)P-labelled ATP or GTP into acid-precipitable material in the absence of exogenous substrates. Their activity was not affected by cyclic AMP or cyclic GMP and was not inhibited by a cyclic nucleotide-dependent protein kinase-inhibitor protein. The kinases are tightly bound to polyribosomes of either origin from infected and non-infected cells, since treatment with 0.5m-NaCl did not dissociate the activity. The enzymes and the enzyme-associated endogenous substrates of cytoplasmic polyribosomes are significantly different from those of the nucleus, and adenovirus infection of the cells did not alter the nature of the enzymes or the substrates at 18-20h after infection. Nuclear kinases catalysed 3-4-fold more phosphate incorporation than did the cytoplasmic kinases. They did not phosphorylate endogenous substrates in the cytoplasmic preparations, and vice versa, which suggests that such substrates for cytoplasmic and nuclear kinases are specific. Polyacrylamide-gel electrophoresis of the phosphorylated proteins revealed the presence of a higher number of endogenous substrates in the nuclear preparation. The nuclear kinases phosphorylated all histones from HeLa cells, but the cytoplasmic ones phosphorylated predominantly the histone of mol.wt. 12000. Bovine heart kinase phosphorylated several low-molecular-weight cytoplasmic proteins and no nuclear proteins. With a DEAE-cellulose column either enzyme activity could be resolved into a number of peaks. The substrate specificities of these peaks indicate that there are at least two different forms of the enzyme in each preparation of polyribosomes.

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Isolation of a distinct pool of polyribosomes from nuclei of adenovirus-infected HeLa cells

Cited by 16 publications

References 21 publications

Differences in sequence content of nuclear and cytoplasmic polyribosomal RNA from adenovirus-infected cells

Differences in sequence content of nuclear and cytoplasmic polyribosomal RNA from adenovirus-infected cells

The path that RNA takes from the nucleus to the cytoplasm: a trip with some surprises

Cyclic nucleotide-independent protein kinases bound to cytoplasmic and nuclear polyribosomes in non-infected and adenovirus-infected HeLa cells

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