BSAP (Pax5)-importin α1 (Rch1) interaction identifies a nuclear localization sequence

Cecilia R. Kovac, Alexander Emelyanov, Mallika Singh, Nasrin Ashouian, Barbara K. Birshtein

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Abstract

BSAP (Pax5) is an essential transcription factor for early B cell and central nervous system development. In later B cell development, BSAP has been implicated in the regulation of 3' Ig enhancers and a number of B cell- specific genes. Previous studies have suggested a role for BSAP-interacting proteins in the regulation of the function of BSAP. Using the yeast two- hybrid system, we identified importin α1 (Rch1) as a BSAP-interacting protein. Importin α proteins have been shown to escort proteins into the nucleus through interaction with a nuclear localization signal (NLS), composed of short stretches of basic amino acids. A predicted NLS in BSAP (NKRKRDE, located at amino acids 195-201 in the central domain) was confirmed to be essential for interaction with importin α1 by the yeast two-hybrid assay. Physical interaction between BSAP and importin α1 was detected in vitro by a glutathione S-transferase (GST) pulldown assay. The NLS sequence in BSAP conferred nuclear localization to green fluorescent protein (GFP)- BSAP fusion proteins. Although the N-terminal paired (DNA-binding) domain of BSAP also conferred nuclear localization when coupled to green fluorescent protein, this domain did not bind to importin α1 in the yeast two-hybrid assay. The NLS sequence in the central domain of BSAP binds to the C-terminal 98-amino acid fragment of importin α1.

Original languageEnglish (US)
Pages (from-to)16752-16757
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number22
DOIs
StatePublished - Jun 2 2000

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Karyopherins
Nuclear Localization Signals
Two-Hybrid System Techniques
Yeast
Assays
B-Lymphocytes
Cells
Protein Sorting Signals
Green Fluorescent Proteins
Proteins
Amino Acids
Basic Amino Acids
Neurology
Glutathione Transferase
Hybrid systems
Transcription Factors
Fusion reactions
Central Nervous System
Genes
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

BSAP (Pax5)-importin α1 (Rch1) interaction identifies a nuclear localization sequence. / Kovac, Cecilia R.; Emelyanov, Alexander; Singh, Mallika; Ashouian, Nasrin; Birshtein, Barbara K.

In: Journal of Biological Chemistry, Vol. 275, No. 22, 02.06.2000, p. 16752-16757.

Research output: Contribution to journalArticle

Kovac, Cecilia R. ; Emelyanov, Alexander ; Singh, Mallika ; Ashouian, Nasrin ; Birshtein, Barbara K. / BSAP (Pax5)-importin α1 (Rch1) interaction identifies a nuclear localization sequence. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 22. pp. 16752-16757.
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