A targeted mutation at the known collagenase cleavage site in mouse type I collagen impairs tissue remodeling.
Degradation of type I collagen, the most abundant collagen, is initiated by collagenase cleavage at a highly conserved site between Gly775 and Ile776 of the alpha 1 (I) chain. Mutations at or around this site render type I collagen resistant to collagenase digestion in vitro. We show here that mice carrying a collagenase-resistant mutant Col1a-1 transgene die late in embryo-genesis, ascribable to overexpression of the transgene, since the same mutation introduced into the endogenous Col1a-1 gene by gene targeting permitted normal development of mutant mice to young adulthood. With increasing age, animals carrying the targeted mutation developed marked fibrosis of the dermis similar to that in human scleroderma. Postpartum involution of the uterus in the mutant mice was also impaired, with persistence of collagenous nodules in the uterine wall. Although type I collagen from the homozygous mutant mice was resistant to cleavage by human or rat fibroblast collagenases at the helical site, only the rat collagenase cleaved collagen trimers at an additional, novel site in the nonhelical N-telopeptide domain. Our results suggest that cleavage by murine collagenase at the N-telopeptide site could account for resorption of type I collagen during embryonic and early adult life. During intense collagen resorption, however, such as in the immediate postpartum uterus and in the dermis later in life, cleavage at the helical site is essential for normal collagen turnover. Thus, type I collagen is degraded by at least two differentially controlled mechanisms involving collagenases with distinct, but overlapping, substrate specificities.
Liu
X
X
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
Wu
H
H
Byrne
M
M
Jeffrey
J
J
Krane
S
S
Jaenisch
R
R
D000595:Amino Acid Sequence; D000818:Animals; D003094:Collagen / Q000502:physiology*; D017364:Collagenases / Q000378:metabolism*; D005260:Female; D005804:Genes, Lethal; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011499:Protein Processing, Post-Translational; D012871:Skin Diseases / Q000235:genetics / Q000473:pathology; D013329:Structure-Activity Relationship
D003094:Collagen; D017364:Collagenases