Carbohydrate-binding protein 35. Levels of transcription and mRNA accumulation in quiescent and proliferating cells

N. Agrwal, J. L. Wang, P. G. Voss

Research output: Contribution to journalArticle

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Abstract

In previous studies, we observed proliferation-dependent expression and nuclear localization of the lectin, designated carbohydrate-binding protein 35 (CBP35), in 3T3 fibroblasts at the polypeptide level by Western blot and immunofluorescence analysis. In the present study, we have compared the expression of the CBP35 gene in quiescent and proliferative 3T3 cells at the levels of (a) accumulated mRNA by Northern blotting and (b) nuclear transcription by run-off assays. When serum-starved, quiescent cultures of 3T3 cells were stimulated by the addition of serum, there was an increase in the nuclear transcription of the CBP35 gene and in the accumulation of its mRNA early (1-3 h) in the activation process, well before the first wave of DNA synthesis. These increases were not dependent on de novo protein synthesis inasmuch as they occurred even in the presence of cycloheximide. There was also an elevated transcription rate and mRNA level in transformed cells when compared to their normal counterparts. Finally, the expression of CBP35 was compared between sparse, proliferating cultures of 3T3 cells and density-inhibited confluent monolayers of the same cells. Although the rate of transcription of the CBP35 gene was approximately the same in the two cultures, there was a higher level of CBP35 mRNA in the dense cells. Thus, it appears that post-transcriptional mechanisms may be involved in the accumulation of mRNA.

Original languageEnglish (US)
Pages (from-to)17236-17242
Number of pages7
JournalJournal of Biological Chemistry
Volume264
Issue number29
StatePublished - 1989
Externally publishedYes

Fingerprint

Galectin 3
Transcription
3T3 Cells
Messenger RNA
Genes
Fibroblasts
Cycloheximide
Serum
Cell culture
Lectins
Northern Blotting
Fluorescent Antibody Technique
Monolayers
Assays
Cell Count
Western Blotting
Chemical activation
Peptides
DNA
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Carbohydrate-binding protein 35. Levels of transcription and mRNA accumulation in quiescent and proliferating cells. / Agrwal, N.; Wang, J. L.; Voss, P. G.

In: Journal of Biological Chemistry, Vol. 264, No. 29, 1989, p. 17236-17242.

Research output: Contribution to journalArticle

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