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Conspicuous accumulation of transcription elongation repressor hrp130/CA150 on the intron-rich Balbiani ring 3 gene

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Abstract

Chromosomal puffs on the polytene chromosomes in the dipteran Chironomus tentans offer the possibility of comparing the appearance of RNA-binding proteins at different transcription sites. We raised a monoclonal antibody that recognized a 130 kDa protein, designated hrp130. Immunocytological analysis of isolated chromosomes showed that hrp130 is heavily accumulated in a specific puff, called Balbiani ring 3; only occasionally is hrp130 abundant in one or two additional puffs on other chromosomes. The immunolabeling was sensitive to RNase treatment, suggesting that hrp130 is associated with nascent ribonucleoproteins. As shown by immunoelectron microscopy hrp130 is distributed along the active BR3 genes. The full sequence of hrp130 was determined by cDNA cloning. The protein comprises 1028 amino acids and contains three WW domains in the N-terminal half and six FF domains in the C-terminal half of the molecule. The protein is conserved from Caenorhabditis elegans to mammals; the human homolog is known as the transcription elongation repressor CA150. We propose that the abundance of hrp130/CA150 in BR3 is connected with the exceptionally high level of splicing in this locus and that hrp130/CA150 adjusts the transcription rate to the numerous splicing events taking place along the gene to ensure proper splicing.

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Acknowledgements

We thank Lars Wieslander for providing us with Chironomus tentans cDNA libraries. The anti-RNA polymerase II antibody was a gift from Arno L. Greenleaf (Duke University Medical Center, Durham, N.C.). We are grateful to Sergej Masich for computer support and to Birgitta Björkroth and Lise-Marie Fjelkestam for technical assistance. This study was supported by the Swedish Research Council, Human Frontier Science Program Organization, Knut and Alice Wallenberg Foundation, Kjell and Märta Beijer Foundation, Ingabritt and Arne Lundberg Foundation and the Karolinska Institute Research Funds.

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  1. Neus Visa

    Present address: Department of Molecular Biology and Functional Genomics, Stockholm University, 10691, Stockholm, Sweden

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, 17177, Stockholm, Sweden

    Xin Sun, Jian Zhao, Karin Kylberg, Teresa Soop, Kevin Palka, Neus Visa, Alla T. Alzhanova-Ericsson & Bertil Daneholt

  2. Center for Genomics and Bioinformatics, Karolinska Institutet, 17177, Stockholm, Sweden

    Erik Sonnhammer

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  1. Xin Sun
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Correspondence to Bertil Daneholt.

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Sun, X., Zhao, J., Kylberg, K. et al. Conspicuous accumulation of transcription elongation repressor hrp130/CA150 on the intron-rich Balbiani ring 3 gene. Chromosoma 113, 244–257 (2004). https://doi.org/10.1007/s00412-004-0314-4

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