Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Feb 1;100(2):514–520. doi: 10.1083/jcb.100.2.514

Developmentally regulated mRNAs in 3T3-adipocytes: analysis of transcriptional control

PMCID: PMC2113435  PMID: 3968175

Abstract

We have investigated the regulation of mRNA synthesis during 3T3- adipocyte differentiation by measuring the transcription of specific genes in isolated preadipocyte and adipocyte nuclei. Transcription was assayed by hybridization of newly synthesized RNA to cDNA clones coding for glycerophosphate dehydrogenase (GPD), the induced protein of 13K which is shown here to be related to myelin protein P-2, the induced protein of 28K, actin, and two RNAs that are not developmentally regulated. Transcription of GPD and 13K was observed in adipocyte but not preadipocyte nuclei. Actin was transcribed in both types of nuclei but at a lower level in adipocytes. For most of the RNAs examined, there was a consistent relationship between amounts of nuclear transcription and the abundance of the corresponding cytoplasmic mRNA in adipocytes. However, 13K and 28K mRNAs are 10-100 times more abundant than would be predicted by their nuclear transcription alone. Preliminary mRNA turnover experiments in which 5,6-dichloro-1-beta-D- ribofuranosylbenzimidazole was used to inhibit mRNA synthesis suggest that these mRNAs are much more stable in the adipocyte cytoplasm than the other mRNAs examined. These results indicate that the transcription of specific genes is increased during adipocyte differentiation and suggest that other levels of control, particularly mRNA stability, may contribute to the relative abundance of certain developmentally- regulated mRNAs in adipocytes.

Full Text

The Full Text of this article is available as a PDF (779.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alpers D. H., Strauss A. W., Ockner R. K., Bass N. M., Gordon J. I. Cloning of a cDNA encoding rat intestinal fatty acid binding protein. Proc Natl Acad Sci U S A. 1984 Jan;81(2):313–317. doi: 10.1073/pnas.81.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berger S. L., Birkenmeier C. S. Inhibition of intractable nucleases with ribonucleoside--vanadyl complexes: isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry. 1979 Nov 13;18(23):5143–5149. doi: 10.1021/bi00590a018. [DOI] [PubMed] [Google Scholar]
  3. Bernlohr D. A., Angus C. W., Lane M. D., Bolanowski M. A., Kelly T. J., Jr Expression of specific mRNAs during adipose differentiation: identification of an mRNA encoding a homologue of myelin P2 protein. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5468–5472. doi: 10.1073/pnas.81.17.5468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Castellot J. J., Jr, Karnovsky M. J., Spiegelman B. M. Differentiation-dependent stimulation of neovascularization and endothelial cell chemotaxis by 3T3 adipocytes. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5597–5601. doi: 10.1073/pnas.79.18.5597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chung S., Landfear S. M., Blumberg D. D., Cohen N. S., Lodish H. F. Synthesis and stability of developmentally regulated dictyostelium mRNAs are affected by cell--cell contact and cAMP. Cell. 1981 Jun;24(3):785–797. doi: 10.1016/0092-8674(81)90104-5. [DOI] [PubMed] [Google Scholar]
  6. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  7. Derman E., Krauter K., Walling L., Weinberger C., Ray M., Darnell J. E., Jr Transcriptional control in the production of liver-specific mRNAs. Cell. 1981 Mar;23(3):731–739. doi: 10.1016/0092-8674(81)90436-0. [DOI] [PubMed] [Google Scholar]
  8. Dreyfuss G., Adam S. A., Choi Y. D. Physical change in cytoplasmic messenger ribonucleoproteins in cells treated with inhibitors of mRNA transcription. Mol Cell Biol. 1984 Mar;4(3):415–423. doi: 10.1128/mcb.4.3.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eriksson U., Sundelin J., Rask L., Peterson P. A. The NH2-terminal amino acid sequence of cellular retinoic-acid binding protein from rat testis. FEBS Lett. 1981 Nov 30;135(1):70–72. doi: 10.1016/0014-5793(81)80945-3. [DOI] [PubMed] [Google Scholar]
  10. Green H., Kehinde O. Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell. 1976 Jan;7(1):105–113. doi: 10.1016/0092-8674(76)90260-9. [DOI] [PubMed] [Google Scholar]
  11. Guyette W. A., Matusik R. J., Rosen J. M. Prolactin-mediated transcriptional and post-transcriptional control of casein gene expression. Cell. 1979 Aug;17(4):1013–1023. doi: 10.1016/0092-8674(79)90340-4. [DOI] [PubMed] [Google Scholar]
  12. Ishaque A., Hofmann T., Eylar E. H. The complete amino acid sequence of the rabbit P2 protein. J Biol Chem. 1982 Jan 25;257(2):592–595. [PubMed] [Google Scholar]
  13. Ishaque A., Szymanska I., Ramwani J., Eylar E. H. Allergic neuritis: phospholipid requirement for the disease-inducing conformation of the P2 protein. Biochim Biophys Acta. 1981 Jun 29;669(1):28–32. doi: 10.1016/0005-2795(81)90219-1. [DOI] [PubMed] [Google Scholar]
  14. Kuri-Harcuch W., Green H. Adipose conversion of 3T3 cells depends on a serum factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6107–6109. doi: 10.1073/pnas.75.12.6107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maizels N. Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976 Nov;9(3):431–438. doi: 10.1016/0092-8674(76)90088-x. [DOI] [PubMed] [Google Scholar]
  16. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  17. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  18. Ockner R. K., Manning J. A., Poppenhausen R. B., Ho W. K. A binding protein for fatty acids in cytosol of intestinal mucosa, liver, myocardium, and other tissues. Science. 1972 Jul 7;177(4043):56–58. doi: 10.1126/science.177.4043.56. [DOI] [PubMed] [Google Scholar]
  19. Ong D. E., Chytil F. Retinoic acid-binding protein in rat tissue. Partial purification and comparison to rat tissue retinol-binding protein. J Biol Chem. 1975 Aug 10;250(15):6113–6117. [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sehgal P. B., Darnell J. E., Jr, Tamm I. The inhibition by DRB (5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole) of hnRNA and mRNA production in HeLa cells. Cell. 1976 Nov;9(3):473–480. doi: 10.1016/0092-8674(76)90092-1. [DOI] [PubMed] [Google Scholar]
  22. Sehgal P. B., Tamm I. Halogenated benzimidazole ribosides, Novel inhibitors of RNA synthesis. Biochem Pharmacol. 1978;27(21):2475–2485. doi: 10.1016/0006-2952(78)90313-1. [DOI] [PubMed] [Google Scholar]
  23. Spiegelman B. M., Farmer S. R. Decreases in tubulin and actin gene expression prior to morphological differentiation of 3T3 adipocytes. Cell. 1982 May;29(1):53–60. doi: 10.1016/0092-8674(82)90089-7. [DOI] [PubMed] [Google Scholar]
  24. Spiegelman B. M., Frank M., Green H. Molecular cloning of mRNA from 3T3 adipocytes. Regulation of mRNA content for glycerophosphate dehydrogenase and other differentiation-dependent proteins during adipocyte development. J Biol Chem. 1983 Aug 25;258(16):10083–10089. [PubMed] [Google Scholar]
  25. Spiegelman B. M., Green H. Control of specific protein biosynthesis during the adipose conversion of 3T3 cells. J Biol Chem. 1980 Sep 25;255(18):8811–8818. [PubMed] [Google Scholar]
  26. Spiegelman B. M., Green H. Cyclic AMP-mediated control of lipogenic enzyme synthesis during adipose differentiation of 3T3 cells. Cell. 1981 May;24(2):503–510. doi: 10.1016/0092-8674(81)90341-x. [DOI] [PubMed] [Google Scholar]
  27. Takahashi K., Odani S., Ono T. A close structural relationship of rat liver Z-protein to cellular retinoid binding proteins and peripheral nerve myelin P2 protein. Biochem Biophys Res Commun. 1982 Jun 30;106(4):1099–1105. doi: 10.1016/0006-291x(82)91225-6. [DOI] [PubMed] [Google Scholar]
  28. Weiss G. H., Rosen O. M., Rubin C. S. Regulation of fatty acid synthetase concentration and activity during adipocyte differentiation. Studies on 3T3-L1 cells. J Biol Chem. 1980 May 25;255(10):4751–4757. [PubMed] [Google Scholar]
  29. White B. A., Bancroft F. C. Cytoplasmic dot hybridization. Simple analysis of relative mRNA levels in multiple small cell or tissue samples. J Biol Chem. 1982 Aug 10;257(15):8569–8572. [PubMed] [Google Scholar]
  30. Wilbur W. J., Lipman D. J. Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci U S A. 1983 Feb;80(3):726–730. doi: 10.1073/pnas.80.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES