Cloning and characterization of mouse cullin4B/E3 ubiquitin ligase

Rachana Tripathi, Seetharama S. Konduru, Satya Keerthi Kota, Usha K. Srinivas

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Heat induced differentiation of mouse embryonal carcinoma cells PCC4 has been reported earlier. We have further characterized the phenotype of the differentiated cells and by DD-RT-PCR identified several partial cDNAs that are differentially expressed during differentiation. Nucleotide homology search revealed that the genes corresponding to some of the up-regulated partial cDNAs are indeed part of differentiation pathway. 5′ extension of an EST that has homology to one of the partial cDNAs led to the identification of mouse cullin4B. Cullin4B is coded by a separate gene and has a unique and longer amino-terminal end with a putative nuclear localization signal sequence (NLS). We have cloned, expressed and raised antibodies against the amino and carboxy-terminal halves of cullin4B. Immuno staining of differentiated PCC4 cells with N-terminal Cul4B antibody showed enhanced expression of Cul4B and its translocation into the nucleus upon differentiation. Transient transfection of a chimeric gene encoding the N-terminal part of Cul4B fused to green fluorescent protein into PCC4 cells revealed that the protein was localized in the nucleus confirming the functional significance of the putative NLS. Since cullins are involved in recognition of specific proteins for degradation, based on the evidence presented here, we hypothesize that cullin4B is probably involved in differentiation specific degradation/modification of nuclear proteins.

Original languageEnglish
Pages (from-to)329-337
Number of pages9
JournalJournal of Biosciences
Volume30
Issue number3
Publication statusPublished - Jun 2005
Externally publishedYes

Fingerprint

ubiquitin-protein ligase
Ubiquitin-Protein Ligases
Cloning
Organism Cloning
molecular cloning
Nuclear Localization Signals
Complementary DNA
Protein Sorting Signals
nuclear localization signals
mice
Genes
signal peptide
Cullin Proteins
Embryonal Carcinoma Stem Cells
Degradation
Gene encoding
Antibodies
Expressed Sequence Tags
cells
Nuclear Proteins

Keywords

  • Cul4B
  • DD-RT-PCR
  • Differential expression
  • Differentiation
  • Nuclear localization sequence
  • PCC4 cells
  • Translocation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Tripathi, R., Konduru, S. S., Kota, S. K., & Srinivas, U. K. (2005). Cloning and characterization of mouse cullin4B/E3 ubiquitin ligase. Journal of Biosciences, 30(3), 329-337.

Cloning and characterization of mouse cullin4B/E3 ubiquitin ligase. / Tripathi, Rachana; Konduru, Seetharama S.; Kota, Satya Keerthi; Srinivas, Usha K.

In: Journal of Biosciences, Vol. 30, No. 3, 06.2005, p. 329-337.

Research output: Contribution to journalArticle

Tripathi, R, Konduru, SS, Kota, SK & Srinivas, UK 2005, 'Cloning and characterization of mouse cullin4B/E3 ubiquitin ligase', Journal of Biosciences, vol. 30, no. 3, pp. 329-337.
Tripathi, Rachana ; Konduru, Seetharama S. ; Kota, Satya Keerthi ; Srinivas, Usha K. / Cloning and characterization of mouse cullin4B/E3 ubiquitin ligase. In: Journal of Biosciences. 2005 ; Vol. 30, No. 3. pp. 329-337.
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