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Identification and characterization of four novel peptide motifs that recognize distinct regions of the transcription factor CP2
Author(s) -
Kang Ho Chul,
Chung Bo Mee,
Chae Ji Hyung,
Yang SungIl,
Kim Chan Gil,
Kim Chul Geun
Publication year - 2005
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2005.04564.x
Subject(s) - transcription factor , yy1 , gene silencing , biology , peptide , binding site , peptide sequence , gene , consensus sequence , microbiology and biotechnology , sequence motif , plasma protein binding , immunoprecipitation , genetics , promoter , biochemistry , gene expression
Although ubiquitously expressed, the transcriptional factor CP2 also exhibits some tissue‐ or stage‐specific activation toward certain genes such as globin in red blood cells and interleukin‐4 in T helper cells. Because this specificity may be achieved by interaction with other proteins, we screened a peptide display library and identified four consensus motifs in numerous CP2‐binding peptides: HXPR, PHL, ASR and PXHXH. Protein‐database searching revealed that RE‐1 silencing factor (REST), Yin‐Yang1 (YY1) and five other proteins have one or two of these CP2‐binding motifs. Glutathione S ‐transferase pull‐down and coimmunoprecipitation assays showed that two HXPR motif‐containing proteins REST and YY1 indeed were able to bind CP2. Importantly, this binding to CP2 was almost abolished when a double amino acid substitution was made on the HXPR sequence of REST and YY1 proteins. The suppressing effect of YY1 on CP2's transcriptional activity was lost by this point mutation on the HXPR sequence of YY1 and reduced by an HXPR‐containing peptide, further supporting the interaction between CP2 and YY1 via the HXPR sequence. Mapping the sites on CP2 for interaction with the four distinct CP2‐binding motifs revealed at least three different regions on CP2. This suggests that CP2 recognizes several distinct binding motifs by virtue of employing different regions, thus being able to interact with and regulate many cellular partners.