Abstract
Incubation of HeLa cells at suboptimal temperature has been used to study the synthesis of 45S ribosomal RNA precursor and the individual steps of the subsequent processing to 28S RNA. Below 20°C no detectable 45S RNA is formed. The processing of 45S RNA to 32S RNA ceases around 15°C, and the processing of 32S RNA to 28S RNA is inhibited near 25°C. Prolonged incubation at reduced temperature results in further modification of the processing, resulting in the apparent accumulation of 41S RNA. The products of these reactions at reduced temperature appear normal in that the ribosomal RNA made at 27°C can be isolated from functional polyribosomes in the cytoplasm after a short incubation at 37°C.
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- Amaldi F., Attardi G. Partial sequence analysis of ribosomal RNA from HeLa cells. I. Oligonucleotide pattern of 28 s and 18 s RNA after pancreatic ribonuclease digestion. J Mol Biol. 1968 May 14;33(3):737–755. doi: 10.1016/0022-2836(68)90317-3. [DOI] [PubMed] [Google Scholar]
- Bishop D. H., Claybrook J. R., Spiegelman S. Electrophoretic separation of viral nucleic acids on polyacrylamide gels. J Mol Biol. 1967 Jun 28;26(3):373–387. doi: 10.1016/0022-2836(67)90310-5. [DOI] [PubMed] [Google Scholar]
- Craig N. C., Perry R. P. Aberrant intranucleolar maturation of ribosomal precursors in the absence of protein synthesis. J Cell Biol. 1970 Jun;45(3):554–564. doi: 10.1083/jcb.45.3.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
- EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
- GIRARD M., PENMAN S., DARNELL J. E. THE EFFECT OF ACTINOMYCIN ON RIBOSOME FORMATION IN HELA CELLS. Proc Natl Acad Sci U S A. 1964 Feb;51:205–211. doi: 10.1073/pnas.51.2.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horisberger M., Amos H. Ribonucleic acid synthesis and transport in animal cells at 27 degrees C. Biochem J. 1970 Apr;117(2):347–353. doi: 10.1042/bj1170347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jeanteur P., Amaldi F., Attardi G. Partial sequence analysis of ribosomal RNA from HeLa cells. II. Evidence for sequences of non-ribosmal type in 45 and 32 s ribosomal RNA precursors. J Mol Biol. 1968 May 14;33(3):757–775. doi: 10.1016/0022-2836(68)90318-5. [DOI] [PubMed] [Google Scholar]
- Levine E. M., Thomas L., McGregor D., Hayflick L., Eagle H. Altered nucleic acid metabolism in human cell cultures infected with mycoplasma. Proc Natl Acad Sci U S A. 1968 Jun;60(2):583–589. doi: 10.1073/pnas.60.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maden B. E., Vaughan M. H., Warner J. R., Darnell J. E. Effects of valine deprivation on ribosome formation in HeLa cells. J Mol Biol. 1969 Oct 28;45(2):265–275. doi: 10.1016/0022-2836(69)90104-1. [DOI] [PubMed] [Google Scholar]
- PERRY R. P. ROLE OF THE NUCLEOLUS IN RIBONUCLEIC ACID METABOLISM AND OTHER CELLULAR PROCESSES. Natl Cancer Inst Monogr. 1964 May;14:73–89. [PubMed] [Google Scholar]
- Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
- Penman S., Scherrer K., Becker Y., Darnell J. E. POLYRIBOSOMES IN NORMAL AND POLIOVIRUS-INFECTED HELA CELLS AND THEIR RELATIONSHIP TO MESSENGER-RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):654–662. doi: 10.1073/pnas.49.5.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
- SCHERRER K., DARNELL J. E. Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun. 1962 Jun 4;7:486–490. doi: 10.1016/0006-291x(62)90341-8. [DOI] [PubMed] [Google Scholar]
- SCHERRER K., LATHAM H., DARNELL J. E. Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc Natl Acad Sci U S A. 1963 Feb 15;49:240–248. doi: 10.1073/pnas.49.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scholtissek C. Nucleotide metabolism in tissue culture cells at low temperatures. I. Phosphorylation of nucleosides and deoxynucleosides in vivo. Biochim Biophys Acta. 1967 Sep 26;145(2):228–237. doi: 10.1016/0005-2787(67)90041-x. [DOI] [PubMed] [Google Scholar]
- Soeiro R., Vaughan M. H., Warner J. R., Darnell J. E., Jr The turnover of nuclear DNA-like RNA in HeLa cells. J Cell Biol. 1968 Oct;39(1):112–118. doi: 10.1083/jcb.39.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vesco C., Penman S. The fractionation of nuclei and the integrity of purified nucleoli in HeLa cells. Biochim Biophys Acta. 1968 Nov 20;169(1):188–195. doi: 10.1016/0005-2787(68)90019-1. [DOI] [PubMed] [Google Scholar]
- Warner J. R., Soeiro R. Nascent ribosomes from HeLa cells. Proc Natl Acad Sci U S A. 1967 Nov;58(5):1984–1990. doi: 10.1073/pnas.58.5.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinberg R. A., Loening U., Willems M., Penman S. Acrylamide gel electrophoresis of HeLa cell nucleolar RNA. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1088–1095. doi: 10.1073/pnas.58.3.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinberg R. A., Penman S. Processing of 45 s nucleolar RNA. J Mol Biol. 1970 Jan 28;47(2):169–178. doi: 10.1016/0022-2836(70)90337-2. [DOI] [PubMed] [Google Scholar]
- Zimmerman E. F., Holler B. W. Methylation of 45 s ribosomal RNA precursor in HeLa cells. J Mol Biol. 1967 Jan 28;23(2):149–161. doi: 10.1016/s0022-2836(67)80023-8. [DOI] [PubMed] [Google Scholar]