Although human nucleoporin Tpr is frequently deregulated in cancer, its roles are poorly understood. Here we show that Tpr depletion generates transcription-dependent replication stress, DNA breaks, and genomic instability. DNA fiber assays and electron microscopy visualization of replication intermediates show that Tpr deficient cells exhibit slow and asymmetric replication forks under replication stress. Tpr deficiency evokes enhanced levels of DNA-RNA hybrids. Additionally, complementary proteomic strategies identify a network of Tpr-interacting proteins mediating RNA processing, such as MATR3 and SUGP2, and functional experiments confirm that their depletion trigger cellular phenotypes shared with Tpr deficiency. Mechanistic studies reveal the interplay of Tpr with GANP, a component of the TREX-2 complex. The Tpr-GANP interaction is supported by their shared protein level alterations in a cohort of ovarian carcinomas. Our results reveal links between nucleoporins, DNA transcription and replication, and the existence of a network physically connecting replication forks with transcription, splicing, and mRNA export machinery.

The human nucleoporin Tpr protects cells from RNA-mediated replication stress / M. Kosar, M. Giannattasio, D. Piccini, A. Maya-Mendoza, F. Garcia-Benitez, J. Bartkova, S.I. Barroso, H. Gaillard, E. Martini, U. Restuccia, M.A. Ramirez-Otero, M. Garre, E. Verga, M. Andujar-Sanchez, S. Maynard, Z. Hodny, V. Costanzo, A. Kumar, A. Bachi, A. Aguilera, J. Bartek, M. Foiani. - In: NATURE COMMUNICATIONS. - ISSN 2041-1723. - 12:1(2021 Jun 24), pp. 3937.1-3937.18. [10.1038/s41467-021-24224-3]

The human nucleoporin Tpr protects cells from RNA-mediated replication stress

M. Giannattasio
Secondo
;
U. Restuccia;V. Costanzo;M. Foiani
Ultimo
2021-06-24

Abstract

Although human nucleoporin Tpr is frequently deregulated in cancer, its roles are poorly understood. Here we show that Tpr depletion generates transcription-dependent replication stress, DNA breaks, and genomic instability. DNA fiber assays and electron microscopy visualization of replication intermediates show that Tpr deficient cells exhibit slow and asymmetric replication forks under replication stress. Tpr deficiency evokes enhanced levels of DNA-RNA hybrids. Additionally, complementary proteomic strategies identify a network of Tpr-interacting proteins mediating RNA processing, such as MATR3 and SUGP2, and functional experiments confirm that their depletion trigger cellular phenotypes shared with Tpr deficiency. Mechanistic studies reveal the interplay of Tpr with GANP, a component of the TREX-2 complex. The Tpr-GANP interaction is supported by their shared protein level alterations in a cohort of ovarian carcinomas. Our results reveal links between nucleoporins, DNA transcription and replication, and the existence of a network physically connecting replication forks with transcription, splicing, and mRNA export machinery.
Acetyltransferases; Cell Survival; DNA Damage; Genomic Instability; HeLa Cells; Humans; Intracellular Signaling Peptides and Proteins; Neoplasms; Nuclear Pore Complex Proteins; Protein Interaction Maps; Proto-Oncogene Proteins; RNA Transport; DNA Replication
Settore BIO/11 - Biologia Molecolare
Settore MED/04 - Patologia Generale
Article (author)
File in questo prodotto:
File Dimensione Formato  
KosaretalNatureCommunication2021.pdf

accesso aperto

Tipologia: Publisher's version/PDF
Dimensione 4.57 MB
Formato Adobe PDF
4.57 MB Adobe PDF Visualizza/Apri
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/906517
Citazioni
  • ???jsp.display-item.citation.pmc??? 2
  • Scopus 2
  • ???jsp.display-item.citation.isi??? 5
social impact