Point mutations within AT-hook domains of the HMGI homologue HMGIYL1 affect binding to gene promoter but not to four-way junction DNA.
High-mobility group I/Y (HMGI/Y) proteins are chromosomal proteins involved in gene and chromatin regulation. Elevated levels of HMGI/Y proteins were reported in diverse malignant tumors, and rearrangements of their genes are casually involved in the development of benign tumors. In humans, the chromosomal locus Xp22 has been often found to be affected in diverse benign mesenchymal tumors. Recent studies revealed that this region contains a retropseudogene HMGIYL1 which potentially can be activated in a way of "exonization" upon aberrations involving this region. The coding sequence of the HMGIY-L1 is highly homologous to the HMGI(Y) gene. On the protein level, both HMGIYL1 and HMGI differ at few amino acid residues, including their putative DNA-binding domains (DBDs). Here we have approached the question of whether the HMGIYL1 product would be able to adopt a role of HMGI in the context of binding to gene promoters and chromatin. Comparative binding studies, employing protein footprinting technique, revealed that HMGIYL1 has lost the ability to bind to the promoter of the interferon beta gene, but retained its high affinity for the four-way junction DNA. Our results stress the importance of particular residues within the DBDs for DNA binding and demonstrate that tight binding of HMGI/Y proteins to the four-way junction DNA can be achieved in alternative ways. The binding of HMGIYL1 to four-way junction DNA suggests that activation of the HMGIYL1 gene would yield a protein sharing some binding properties with HMG1-box proteins and histone H1. Thus, the HMGIYL1 could interplay together with these components in chromatin regulation.
Schwanbeck
R
R
Zoologisches Institut-Entwicklungsbiologie, Universität Göttingen, D-37073 Göttingen, Germany.
Gerharz
M
M
Drung
A
A
Rogalla
P
P
Piekielko
A
A
Blank
C
C
Bullerdiek
J
J
Wiśniewski
J R
JR
D021581:Active Transport, Cell Nucleus / Q000235:genetics; D000225:Adenine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D002877:Chromosomes, Human / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D006609:High Mobility Group Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D016899:Interferon-beta / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D023302:Protein Footprinting; D017434:Protein Structure, Tertiary / Q000235:genetics; D011544:Pseudogenes; D017386:Sequence Homology, Amino Acid; D013941:Thymine / Q000378:metabolism
D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D008164:Luminescent Proteins; D010750:Phosphoproteins; D049452:Green Fluorescent Proteins; D016899:Interferon-beta; D000225:Adenine; D013941:Thymine