Mid-region parathyroid hormone-related protein (PTHrP) binds chromatin of MDA-MB231 breast cancer cells and isolated oligonucleotides "in vitro".
We have previously shown that PTHrP(38-94)-amide restrains growth and invasion "in vitro", causes striking toxicity and accelerates death of some breast cancer cell lines, the most responsive being MDA-MB231 whose tumorigenesis was also attenuated "in vivo". PTHrP(38-94)-amide contains the domain implicated in the nuclear import of PTHrP. Although the nucleus was identified as a destination for mid-region PTHrP, evidence for direct DNA-binding capability is lacking to date. Here, we examined the localization of PTHrP(38-94)-amide within MDA-MB231 cells and within metaphase spread preparations and characterized its DNA-binding properties, employing a combination of immunocytochemical, cytogenetic, "whole genome"/conventional PCR, EMSA and DNase footprinting techniques. The results obtained: (i) show that PTHrP(38-94)-amide gains access to the nuclear compartment of MDA-MB231 cell; (ii) demonstrate that PTHrP(38-94)-amide is a DNA-binding peptide; and, (iii) represent the first data to date on the potential molecular targets in both cellular chromatin and isolated oligonucleotides "in vitro".
Sirchia
Rosalia
R
Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Palermo, Viale delle Scienze, 90128, Palermo, Italy.
Priulla
Marcella
M
Sciandrello
Giulia
G
Caradonna
Fabio
F
Barbata
Giuseppa
G
Luparello
Claudio
C
D021581:Active Transport, Cell Nucleus; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000737:chemistry; D016678:Genome; D006801:Humans; D008280:Magnetics; D008677:Metaphase; D008954:Models, Biological; D009841:Oligonucleotides / Q000737:chemistry; D044162:Parathyroid Hormone-Related Protein / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding
D002843:Chromatin; D009841:Oligonucleotides; D044162:Parathyroid Hormone-Related Protein; D010455:Peptides; D004247:DNA