The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins

Autor: Belinda J. Westman, Kate G. R. Quinlan, Nicholas E. Shepherd, Merlin Crossley, Molly M.K. Clifton, Sock Yue Thong, Mitchell R. O’Connell, Joel P. Mackay, Michael W. Webster, Gerd A. Blobel
Rok vydání: 2014
Předmět:
Models
Molecular
Protein Structure
EGF-like domain
Science
Protein domain
Molecular Sequence Data
Gene Expression
Biology
Spectrum analysis techniques
Biochemistry
HAMP domain
Mice
Protein structure
NMR spectroscopy
Protein Domains
Cell Line
Tumor
DNA-binding proteins
Escherichia coli
Animals
Humans
Protein Isoforms
Amino Acid Sequence
Conserved Sequence
Genetics
Multidisciplinary
Sequence Homology
Amino Acid
Biology and life sciences
DHR1 domain
Nuclear Proteins
Proteins
S-Adenosylhomocysteine
Recombinant Proteins
Cell biology
Protein Structure
Tertiary
Regulatory Proteins
Research and analysis methods
Cyclic nucleotide-binding domain
Histone methyltransferase
Medicine
Protein Structure Determination
Sequence Alignment
Transcription Activators
Binding domain
Transcription Factors
Research Article
Zdroj: PLoS ONE
PLoS ONE, Vol 9, Iss 8, p e106011 (2014)
ISSN: 1932-6203
Popis: FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being.
Databáze: OpenAIRE
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