HMG box 4 is the principal determinant of species specificity in the RNA polymerase I transcription factor UBF.
Transcription of ribosomal genes requires, in addition to RNA polymerase I, the trans-acting factors UBF and Rib1 in Xenopus or SL1 in humans. RNA polymerase I transcription is remarkably species specific. Between closely related species SL1 is the sole determinant of this specificity. Between more distantly related species, however, UBF is also a component of this species specificity. Xenopus UBF cannot function in human RNA polymerase I transcription and human UBF cannot function in Xenopus RNA polymerase I transcription. Xenopus and human UBFs are remarkably similar at the amino acid sequence level, both containing multiple HMG box DNA binding motifs. The only major difference between xUBF and hUBF is the lack of a HMG box 4 equivalent in xUBF. Utilizing a series of hybrid UBF molecules we have identified HMG box 4 as the principal determinant of species specificity. Addition of human HMG box 4 to xUBF converts it to a form that functions in human RNA polymerase I transcription. Deletion of HMG box 4 from hUBF converts it to a form that functions in Xenopus RNA polymerase I transcription. Furthermore, mutations within Xenopus UBF demonstrate that UBF requires a precise arrangement and number of HMG boxes to function in RNA polymerase I transcription.
Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, UK.
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Cairns
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McStay
D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D010957:Plasmids; D035463:Pol1 Transcription Initiation Complex Proteins; D016133:Polymerase Chain Reaction; D012318:RNA Polymerase I / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013045:Species Specificity; D018411:Spodoptera; D014157:Transcription Factors / Q000096:biosynthesis / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D014981:Xenopus
D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D035463:Pol1 Transcription Initiation Complex Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; C063844:transcription factor UBF; D012318:RNA Polymerase I