PUM2, a novel murine puf protein, and its consensus RNA-binding site.

Members of the Puf family of RNA-binding proteins from Drosophila, Caenorhabditis elegans, and Dictyostelium are known to function as translational repressors. To identify mammalian proteins that might regulate posttranscriptional gene expression, we have characterized a novel murine Puf protein, PUM2. Pum2 transcripts were expressed in all murine tissues examined, suggesting the gene influences processes common to many cell types. Like all Puf family members, PUM2 contains a C-terminal RNA-binding domain related to the Drosophila Pumilio homology domain (PUM-HD). Two features found in the amino-terminus of PUM2, regions rich in serine and glutamine/alanine-rich regions, were also identified in most Puf family members. RNA sequences capable of binding with high affinity (6.5 nM) to a 48-kDa recombinant protein containing the PUM2 PUM-HD were isolated by using an iterative amplification-selection protocol (SELEX). The consensus sequence [UGUANAUARNNNNBBBBSCCS] of the PUM2 binding element (PBE) is related to, but distinct from, the 3' end of the Drosophila Nanos response element. The characterization of PUM2 and potential RNA-binding site will assist efforts to assess the extent and mechanism by which mammalian genes are regulated at a posttranscriptional level.

[1]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[2]  R. Klausner,et al.  Regulating the fate of mRNA: The control of cellular iron metabolism , 1993, Cell.

[3]  D. Schümperli,et al.  Faithful cell-cycle regulation of a recombinant mouse histone H4 gene is controlled by sequences in the 3'-terminal part of the gene. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Masashi Yamada,et al.  Maternal Pumilio acts together with Nanos in germline development in Drosophila embryos , 1999, Nature Cell Biology.

[5]  R. Lehmann,et al.  Involvement of the pumilio gene in the transport of an abdominal signal in the Drosophila embryo , 1987, Nature.

[6]  A. Spradling,et al.  A novel group of pumilio mutations affects the asymmetric division of germline stem cells in the Drosophila ovary. , 1997, Development.

[7]  L. Warnock,et al.  The 3' untranslated region of IL-1beta regulates protein production. , 1997, Journal of immunology.

[8]  Myriam Gorospe,et al.  HuR regulates cyclin A and cyclin B1 mRNA stability during cell proliferation , 2000, The EMBO journal.

[9]  J. Richter,et al.  The control of cyclin B1 mRNA translation during mouse oocyte maturation. , 2000, Developmental biology.

[10]  M. Parisi,et al.  The Drosophila pumilio gene encodes two functional protein isoforms that play multiple roles in germline development, gonadogenesis, oogenesis and embryogenesis. , 1999, Genetics.

[11]  J. Keene,et al.  Embryonic lethal abnormal visual RNA-binding proteins involved in growth, differentiation, and posttranscriptional gene expression. , 1997, American journal of human genetics.

[12]  R. Lehmann,et al.  Pumilio is essential for function but not for distribution of the Drosophila abdominal determinant Nanos. , 1992, Genes & development.

[13]  M. Whitfield,et al.  The protein that binds the 3' end of histone mRNA: a novel RNA-binding protein required for histone pre-mRNA processing. , 1996, Genes & development.

[14]  I. Herskowitz,et al.  Post‐transcriptional regulation through the HO 3′‐UTR by Mpt5, a yeast homolog of Pumilio and FBF , 2001, The EMBO journal.

[15]  R. Wharton,et al.  The Pumilio RNA-binding domain is also a translational regulator. , 1998, Molecular cell.

[16]  N. Nomura,et al.  Prediction of the coding sequences of unidentified human genes. VI. The coding sequences of 80 new genes (KIAA0201-KIAA0280) deduced by analysis of cDNA clones from cell line KG-1 and brain. , 1996, DNA research : an international journal for rapid publication of reports on genes and genomes.

[17]  P. Zamore,et al.  Crystal structure of a Pumilio homology domain. , 2001, Molecular cell.

[18]  Yi-shuian Huang,et al.  CPEB, Maskin, and Cyclin B1 mRNA at the Mitotic Apparatus Implications for Local Translational Control of Cell Division , 2000, Cell.

[19]  Y Nagahama,et al.  Biochemical Identification of Xenopus Pumilio as a Sequence-specific Cyclin B1 mRNA-binding Protein That Physically Interacts with a Nanos Homolog, Xcat-2, and a Cytoplasmic Polyadenylation Element-binding Protein* , 2001, The Journal of Biological Chemistry.

[20]  H. Ruley,et al.  Fus deficiency in mice results in defective B-lymphocyte development and activation, high levels of chromosomal instability and perinatal death , 2000, Nature Genetics.

[21]  R. Wharton,et al.  Recruitment of Nanos to hunchback mRNA by Pumilio. , 1999, Genes & development.

[22]  R. Lehmann,et al.  Nanos and Pumilio have critical roles in the development and function of Drosophila germline stem cells. , 1998, Development.

[23]  G. Shaw,et al.  A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation , 1986, Cell.

[24]  The PUMILIO-RNA interaction: a single RNA-binding domain monomer recognizes a bipartite target sequence. , 1999, Biochemistry.

[25]  D. Richter,et al.  Identification of a cis-Acting Dendritic Targeting Element in MAP2 mRNAs , 1999, The Journal of Neuroscience.

[26]  Stanley Fields,et al.  A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line , 1997, Nature.

[27]  A. Aggarwal,et al.  Structure of Pumilio Reveals Similarity between RNA and Peptide Binding Motifs , 2001, Cell.

[28]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[29]  G. Struhl,et al.  RNA regulatory elements mediate control of Drosophila body pattern by the posterior morphogen nanos , 1991, Cell.

[30]  D. Glover,et al.  3' non-translated sequences in Drosophila cyclin B transcripts direct posterior pole accumulation late in oogenesis and peri-nuclear association in syncytial embryos. , 1992, Development.

[31]  O. Uhlenbeck,et al.  Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. , 1987, Nucleic acids research.

[32]  M. Suyama,et al.  HUGE: a database for human large proteins identified in the Kazusa cDNA sequencing project. , 2000, Nucleic acids research.

[33]  Marvin Wickens,et al.  NANOS-3 and FBF proteins physically interact to control the sperm–oocyte switch in Caenorhabditis elegans , 1999, Current Biology.

[34]  J. Richter,et al.  Cytoplasmic polyadenylation elements mediate masking and unmasking of cyclin B1 mRNA , 1999, The EMBO journal.

[35]  R. Lehmann,et al.  The Pumilio protein binds RNA through a conserved domain that defines a new class of RNA-binding proteins. , 1997, RNA.

[36]  R. Wharton,et al.  Binding of pumilio to maternal hunchback mRNA is required for posterior patterning in drosophila embryos , 1995, Cell.

[37]  U. Atasoy,et al.  ELAV protein HuA (HuR) can redistribute between nucleus and cytoplasm and is upregulated during serum stimulation and T cell activation. , 1998, Journal of cell science.

[38]  R. Wharton,et al.  Drosophila Brain Tumor is a translational repressor. , 2001, Genes & development.

[39]  C. Thompson,et al.  An inducible cytoplasmic factor (AU-B) binds selectively to AUUUA multimers in the 3' untranslated region of lymphokine mRNA , 1991, Molecular and cellular biology.

[40]  David J. States,et al.  Identification of protein coding regions by database similarity search , 1993, Nature Genetics.

[41]  A. Kuspa,et al.  Starvation promotes Dictyostelium development by relieving PufA inhibition of PKA translation through the YakA kinase pathway. , 1999, Development.

[42]  P. Macdonald,et al.  The Drosophila pumilio gene: an unusually long transcription unit and an unusual protein. , 1992, Development.

[43]  J. Keene,et al.  ELAV tumor antigen, Hel-N1, increases translation of neurofilament M mRNA and induces formation of neurites in human teratocarcinoma cells. , 1999, Genes & development.