Recombinant hnRNP protein A1 and its N-terminal domain show preferential affinity for oligodeoxynucleotides homologous to intron/exon acceptor sites (original) (raw)

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Istituto di Genetica Biochimica ed Evoluzionistica

CNR Via Abbiategrasso 207, 27100 Pavia, Italy

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Istituto di Genetica Biochimica ed Evoluzionistica

CNR Via Abbiategrasso 207, 27100 Pavia, Italy

Search for other works by this author on:

,

Istituto di Genetica Biochimica ed Evoluzionistica

CNR Via Abbiategrasso 207, 27100 Pavia, Italy

Search for other works by this author on:

Istituto di Genetica Biochimica ed Evoluzionistica

CNR Via Abbiategrasso 207, 27100 Pavia, Italy

*To whom correspondence should be addressed

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Revision received:

11 October 1990

Accepted:

11 October 1990

Published:

25 November 1990

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Massimo Buvoli, Fabio Cobianchi, Giuseppe Biamonti, Silvano Riva, Recombinant hnRNP protein A1 and its N-terminal domain show preferential affinity for oligodeoxynucleotides homologous to intron/exon acceptor sites, Nucleic Acids Research, Volume 18, Issue 22, 25 November 1990, Pages 6595–6600, https://doi.org/10.1093/nar/18.22.6595
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Abstract

The reported binding preference of human hnRNP protein A1 for the 3′-splice site of some introns (Swanson and Dreyfuss (1988) EMBO J. 7,3519–3529; Mayrand and Pederson (1990) Nucleic Acids Res. 18, 3307–3318) was tested by assaying in vitro the binding of purified recombinant A1 protein (expressed in bacteria) to synthetic oligodeoxynucleotides (21-mers) of suitable sequence. In such a minimal system we find preferential binding of protein A1 to ollgodeoxy-nucleotide sequences corresponding to the 3′-splice site of IVS1 of human β-globin pre-mRNA and of IVS1 of Adenovirus type 2 major late transcript. Mutation studies demonstrate that the binding specificity is dependent on the known critical domains of this intron region, the AG splice site dinucleotide and polypyrimidine tract, and resides entirely in the short oligonucleotlde sequence. Moreover specific binding does not require the presence of other hnRNP proteins or of snRNP particles. Studies with a truncated recombinant protein demonstrated that the minimal protein sequence determinants for A1 recognition of 3′-splice acceptor site reside entirely in the N-terminal 195 aa of the unmodified protein.

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© 1990 Oxford University Press

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