Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.

ERH is a small, highly evolutionarily conserved nuclear protein of unknown function. Its three-dimensional structure is absolutely unique and it can form a homodimer through a β sheet surface. ERH has been shown to interact, among others, with PDIP46/SKAR and Ciz1. When coexpressed with the latter p...

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Autores principales:	Monika I Banko, Marek K Krzyzanowski, Paulina Turcza, Zuzanna Maniecka, Marta Kulis, Piotr Kozlowski
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c938a38067844c67806b26b6141b2763
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spelling	oai:doaj.org-article:c938a38067844c67806b26b6141b27632021-11-18T08:57:48ZIdentification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.1932-620310.1371/journal.pone.0074885https://doaj.org/article/c938a38067844c67806b26b6141b27632013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24015320/?tool=EBIhttps://doaj.org/toc/1932-6203ERH is a small, highly evolutionarily conserved nuclear protein of unknown function. Its three-dimensional structure is absolutely unique and it can form a homodimer through a β sheet surface. ERH has been shown to interact, among others, with PDIP46/SKAR and Ciz1. When coexpressed with the latter protein, ERH accumulates in replication foci in the nucleus of HeLa cells. Here, we report that when ERH is coexpressed with PDIP46/SKAR in HeLa cells, it is recruited to nuclear speckles, and identify amino acid residues critical for targeting ERH to both these subnuclear structures. ERH H3A Q9A shows a diminished recruitment to nuclear speckles but it is recruited to replication foci. ERH E37A T51A is very poorly recruited to replication foci while still accumulating in nuclear speckles. Consequently, ERH H3A Q9A E37A T51A is recruited neither to nuclear speckles nor to replication foci. The lack of interactions of these three ERH forms with PDIP46/SKAR and/or Ciz1 was further confirmed in vitro by GST pull-down assay. The residues whose substitutions interfere with the accumulation in nuclear speckles are situated on the β sheet surface of ERH, indicating that only the monomer of ERH can interact with PDIP46/SKAR. Substitutions affecting the recruitment to replication foci map to the other side of ERH, near a long loop between the α1 and α2 helices, thus both the monomer and the dimer of ERH could interact with Ciz1. The construction of the ERH mutants not recruited to nuclear speckles or replication foci will facilitate further studies on ERH actions in these subnuclear structures.Monika I BankoMarek K KrzyzanowskiPaulina TurczaZuzanna ManieckaMarta KulisPiotr KozlowskiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e74885 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Monika I Banko Marek K Krzyzanowski Paulina Turcza Zuzanna Maniecka Marta Kulis Piotr Kozlowski Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
description	ERH is a small, highly evolutionarily conserved nuclear protein of unknown function. Its three-dimensional structure is absolutely unique and it can form a homodimer through a β sheet surface. ERH has been shown to interact, among others, with PDIP46/SKAR and Ciz1. When coexpressed with the latter protein, ERH accumulates in replication foci in the nucleus of HeLa cells. Here, we report that when ERH is coexpressed with PDIP46/SKAR in HeLa cells, it is recruited to nuclear speckles, and identify amino acid residues critical for targeting ERH to both these subnuclear structures. ERH H3A Q9A shows a diminished recruitment to nuclear speckles but it is recruited to replication foci. ERH E37A T51A is very poorly recruited to replication foci while still accumulating in nuclear speckles. Consequently, ERH H3A Q9A E37A T51A is recruited neither to nuclear speckles nor to replication foci. The lack of interactions of these three ERH forms with PDIP46/SKAR and/or Ciz1 was further confirmed in vitro by GST pull-down assay. The residues whose substitutions interfere with the accumulation in nuclear speckles are situated on the β sheet surface of ERH, indicating that only the monomer of ERH can interact with PDIP46/SKAR. Substitutions affecting the recruitment to replication foci map to the other side of ERH, near a long loop between the α1 and α2 helices, thus both the monomer and the dimer of ERH could interact with Ciz1. The construction of the ERH mutants not recruited to nuclear speckles or replication foci will facilitate further studies on ERH actions in these subnuclear structures.
format	article
author	Monika I Banko Marek K Krzyzanowski Paulina Turcza Zuzanna Maniecka Marta Kulis Piotr Kozlowski
author_facet	Monika I Banko Marek K Krzyzanowski Paulina Turcza Zuzanna Maniecka Marta Kulis Piotr Kozlowski
author_sort	Monika I Banko
title	Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
title_short	Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
title_full	Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
title_fullStr	Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
title_full_unstemmed	Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.
title_sort	identification of amino acid residues of erh required for its recruitment to nuclear speckles and replication foci in hela cells.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/c938a38067844c67806b26b6141b2763
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Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells.

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