Characterization of the DNA-binding domain of the avian Y-box protein, chkYB-2, and mutational analysis of its single-strand binding motif in the rous sarcoma virus enhancer

Ashok Nambiar, S. K. Swamynathan, Jagannadha C. Kandala, Ramareddy Guntaka

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

chkYB-2 is a sequence-specific, single-stranded DNA binding chicken Y- box protein that promotes Rous sarcoma virus long terminal repeat (RSV LTR)- driven transcription in avian fibroblasts. The DNA-binding domain of chkYB-2 has been mapped by characterizing the DNA binding properties of purified recombinant chkYB-2 mutant polypeptides. The data indicate that the invariant cold shock domain (CSD) is necessary but not sufficient for association with DNA and suggest that another conserved region, adjacent to the carboxyl boundary of the CSD, plays a role in high-affinity DNA binding. chkYB-2 binds to a tandem repeat of the 5'-GTACCACC-3' motif on the RSV LTR. Mutational analysis of this recognition sequence revealed the requirement of an essentially unaltered template for both high-affinity binding by chkYB-2 as well as maximal transcriptional activity of the RSV LTR in vivo. The single- stranded DNA binding activity of chkYB-2 is augmented by Mg2+. The possible significance of this finding for transactivation by a single-strand DNA binding protein is discussed.

Original languageEnglish (US)
Pages (from-to)900-909
Number of pages10
JournalJournal of Virology
Volume72
Issue number2
StatePublished - Feb 1998
Externally publishedYes

Fingerprint

Rous sarcoma virus
terminal repeat sequences
DNA-binding domains
Terminal Repeat Sequences
single-stranded DNA
cold stress
Single-Stranded DNA
DNA
Shock
Proteins
DNA-binding proteins
tandem repeat sequences
proteins
binding properties
transcriptional activation
Tandem Repeat Sequences
fibroblasts
polypeptides
DNA-Binding Proteins
transcription (genetics)

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

Characterization of the DNA-binding domain of the avian Y-box protein, chkYB-2, and mutational analysis of its single-strand binding motif in the rous sarcoma virus enhancer. / Nambiar, Ashok; Swamynathan, S. K.; Kandala, Jagannadha C.; Guntaka, Ramareddy.

In: Journal of Virology, Vol. 72, No. 2, 02.1998, p. 900-909.

Research output: Contribution to journalArticle

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abstract = "chkYB-2 is a sequence-specific, single-stranded DNA binding chicken Y- box protein that promotes Rous sarcoma virus long terminal repeat (RSV LTR)- driven transcription in avian fibroblasts. The DNA-binding domain of chkYB-2 has been mapped by characterizing the DNA binding properties of purified recombinant chkYB-2 mutant polypeptides. The data indicate that the invariant cold shock domain (CSD) is necessary but not sufficient for association with DNA and suggest that another conserved region, adjacent to the carboxyl boundary of the CSD, plays a role in high-affinity DNA binding. chkYB-2 binds to a tandem repeat of the 5'-GTACCACC-3' motif on the RSV LTR. Mutational analysis of this recognition sequence revealed the requirement of an essentially unaltered template for both high-affinity binding by chkYB-2 as well as maximal transcriptional activity of the RSV LTR in vivo. The single- stranded DNA binding activity of chkYB-2 is augmented by Mg2+. The possible significance of this finding for transactivation by a single-strand DNA binding protein is discussed.",
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AB - chkYB-2 is a sequence-specific, single-stranded DNA binding chicken Y- box protein that promotes Rous sarcoma virus long terminal repeat (RSV LTR)- driven transcription in avian fibroblasts. The DNA-binding domain of chkYB-2 has been mapped by characterizing the DNA binding properties of purified recombinant chkYB-2 mutant polypeptides. The data indicate that the invariant cold shock domain (CSD) is necessary but not sufficient for association with DNA and suggest that another conserved region, adjacent to the carboxyl boundary of the CSD, plays a role in high-affinity DNA binding. chkYB-2 binds to a tandem repeat of the 5'-GTACCACC-3' motif on the RSV LTR. Mutational analysis of this recognition sequence revealed the requirement of an essentially unaltered template for both high-affinity binding by chkYB-2 as well as maximal transcriptional activity of the RSV LTR in vivo. The single- stranded DNA binding activity of chkYB-2 is augmented by Mg2+. The possible significance of this finding for transactivation by a single-strand DNA binding protein is discussed.

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