title: Cystatin D locates in the nucleus at sites of active transcription and modulates gene and protein expression.
abstract: Cystatin D is an inhibitor of lysosomal and secreted cysteine proteases. Strikingly, cystatin D has been found to inhibit proliferation, migration, and invasion of colon carcinoma cells indicating tumor suppressor activity that is unrelated to protease inhibition. Here, we demonstrate that a proportion of cystatin D locates within the cell nucleus at specific transcriptionally active chromatin sites. Consistently, transcriptomic analysis show that cystatin D alters gene expression, including that of genes encoding transcription factors such as RUNX1, RUNX2, and MEF2C in HCT116 cells. In concordance with transcriptomic data, quantitative proteomic analysis identified 292 proteins differentially expressed in cystatin D-expressing cells involved in cell adhesion, cytoskeleton, and RNA synthesis and processing. Furthermore, using cytokine arrays we found that cystatin D reduces the secretion of several protumor cytokines such as fibroblast growth factor-4, CX3CL1/fractalkine, neurotrophin 4 oncostatin-M, pulmonary and activation-regulated chemokine/CCL18, and transforming growth factor B3. These results support an unanticipated role of cystatin D in the cell nucleus, controlling the transcription of specific genes involved in crucial cellular functions, which may mediate its protective action in colon cancer.
authors.affiliation: From the Instituto de Investigaciones Biomédicas "Alberto Sols," Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, E-28029 Madrid.
authors.forename: Gemma
authors.initials: G
authors.lastname: Ferrer-Mayorga
authors.affiliation: From the Instituto de Investigaciones Biomédicas "Alberto Sols," Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, E-28029 Madrid.
authors.forename: Silvia
authors.initials: S
authors.lastname: Alvarez-Díaz
authors.affiliation: From the Instituto de Investigaciones Biomédicas "Alberto Sols," Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, E-28029 Madrid.
authors.forename: Noelia
authors.initials: N
authors.lastname: Valle
authors.affiliation: the Centro de Investigación del Cáncer, Consejo Superior de Investigaciones Científicas-Universidad de Salamanca, E-37007 Salamanca.
authors.forename: Javier
authors.initials: J
authors.lastname: De Las Rivas
authors.affiliation: the Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, E-28040 Madrid.
authors.forename: Marta
authors.initials: M
authors.lastname: Mendes
authors.affiliation: the Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, E-28040 Madrid.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Barderas
authors.affiliation: the Proteomics Laboratory, Vall d'Hebron Institute of Oncology, Vall d'Hebron University Hospital, Barcelona, E-08035, and.
authors.forename: Francesc
authors.initials: F
authors.lastname: Canals
authors.affiliation: the Departamento de Anatomía y Biología Celular, Facultad de Medicina, Universidad de Cantabria-Instituto de Investigación Valdecilla, E-39011 Santander, Spain.
authors.forename: Olga
authors.initials: O
authors.lastname: Tapia
authors.affiliation: the Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, E-28040 Madrid.
authors.forename: J Ignacio
authors.initials: JI
authors.lastname: Casal
authors.affiliation: the Departamento de Anatomía y Biología Celular, Facultad de Medicina, Universidad de Cantabria-Instituto de Investigación Valdecilla, E-39011 Santander, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Lafarga
authors.affiliation: From the Instituto de Investigaciones Biomédicas "Alberto Sols," Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, E-28029 Madrid, amunoz@iib.uam.es.
authors.forename: Alberto
authors.initials: A
authors.lastname: Muñoz
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics / Q000378:metabolism; D015891:Cystatins / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D014158:Transcription, Genetic
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D050676:Core Binding Factor Alpha 2 Subunit; D015891:Cystatins; D016207:Cytokines; D064326:MEF2 Transcription Factors; C578040:MEF2C protein, human; D009363:Neoplasm Proteins; C493728:RUNX1 protein, human; C493709:RUNX2 protein, human; C071378:CST5 protein, human
keywords: colon cancer; cysteine protease; cytokine; gene expression; protease inhibitor