title: Strain response in fibroblasts indicates a possible role of the Ca(2+)-dependent nuclear transcription factor NM1 in RNA synthesis.
abstract: On the mechanistic level, response of periodontal fibroblasts permanently exposed to mechanical strain forces in vivo still lacks in clarity. Therefore, we first investigated putative strain modulation of proteins by combined 1D gel electrophoresis-based protein profiling and electrospray tandem mass spectrometry (ESI-MS). Thereafter, the exponential-modified protein abundance index (emPAI) identified strain modulation of cytoskeleton-associated molecules, including decrease in talin and microtubule-associated protein 4 (MAP4), and significant increase in myosin IC (Myo IC), the latter ones regulated by Ca(2+). These findings were corroborated by western blotting (WB) and indirect immunofluorescence (IIF). Regarding the dual function of Myo IC as actin-based cytoplasmic motor protein and nuclear transcription factor NM1, WB and IIF revealed inverse correlation for Myo IC and NM1. During strain application, cytoplasmic increase of Myo IC was counteracted by nuclear NM1 deprivation, the latter coinciding with a decline in RNA quantity. Independent on strain, cytoplasmic Myo IC and nuclear NM1 abundance could be abrogated by the Ca(2+) channel blocker nifedipine, suggesting Ca(2+) dependency of cytoplasmic and/or nuclear Myo IC/NM1 expression. Mechanistically, we conclude that, application of strain appears as causative for the decline in RNA by impacting NM1, thereby indicating the possible role of NM1 in RNA synthesis.
authors.affiliation: Department of Oral Biotechnology, Dental School, University Hospital Freiburg, Hugstetterstrasse 55, 79106 Freiburg, Germany. thorsten.steinberg@uniklinik-freiburg.de
authors.forename: Thorsten
authors.initials: T
authors.lastname: Steinberg
authors.forename: Nelli
authors.initials: N
authors.lastname: Ziegler
authors.forename: Angel
authors.initials: A
authors.lastname: Alonso
authors.forename: Annette
authors.initials: A
authors.lastname: Kohl
authors.forename: Eva
authors.initials: E
authors.lastname: Müssig
authors.forename: Susanne
authors.initials: S
authors.lastname: Proksch
authors.forename: Simon
authors.initials: S
authors.lastname: Schulz
authors.forename: Pascal
authors.initials: P
authors.lastname: Tomakidi
mesh_terms: D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D024461:Myosin Type I / Q000378:metabolism / Q000502:physiology*; D009543:Nifedipine / Q000494:pharmacology; D020543:Proteome / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis*; D021241:Spectrometry, Mass, Electrospray Ionization; D013314:Stress, Mechanical; D016608:Talin / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*
chemical_list: D008869:Microtubule-Associated Proteins; D020543:Proteome; D016608:Talin; D014157:Transcription Factors; D012313:RNA; D024461:Myosin Type I; C460367:MYO1C protein, human; D009543:Nifedipine; D002118:Calcium