Downregulation of collagen synthesis in fibroblasts within three-dimensional collagen lattices involves transcriptional and posttranscriptional mechanisms.
Culturing human fibroblasts in a three-dimensional collagen matrix leads to a reduction of collagen I by more than 90%, both on the level of mRNA steady-state as well as protein. In order to differentiate changes in de novo transcription and posttranscriptional control, nuclear run on assays and pulse/chase experiments determining mRNA stability were used. Our results indicate that de novo transcription of the COL1A1 gene and pro-alpha 1 (I)collagen mRNA half-life are both decreased by 50% in fibroblasts grown in three-dimensional collagen lattices as compared to monolayer cultures. The extracellular matrix therefore elicits signals which are transduced from the cell surface to the inside of fibroblasts resulting in a specific reprogramming of transcriptional as well as posttranscriptional processes.
Eckes
B
B
Department of Dermatology, University of Cologne, Germany.
Mauch
C
C
Hüppe
G
G
Krieg
T
T
D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005782:Gels; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D066298:In Vitro Techniques; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D005782:Gels; D012333:RNA, Messenger; D003094:Collagen