Type I collagen triplet duplication mutation in lethal osteogenesis imperfecta shifts register of α chains throughout the helix and disrupts incorporation of mutant helices into fibrils and extracellular matrix

Wayne A. Cabral, Marianna V. Mertts, Elena Makareeva, Alain Colige, Mustafa Tekin, Arti Pandya, Sergey Leikin, Joan C. Marini

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The majority of collagen mutations causing osteogenesis imperfecta (OI) are glycine substitutions that disrupt formation of the triple helix. A rare type of collagen mutation consists of a duplication or deletion of one or two Gly-X-Y triplets. These mutations shift the register of collagen chains with respect to each other in the helix but do not interrupt the triplet sequence, yet they have severe clinical consequences. We investigated the effect of shifting the register of the collagen helix by a single Gly-X-Y triplet on collagen assembly, stability, and incorporation into fibrils and matrix. These studies utilized a triplet duplication in COL1A1 exon 44 that occurred in the cDNA and gDNA of two siblings with lethal OI. The normal allele encodes three identical Gly-Ala-Hyp triplets at aa 868-876, whereas the mutant allele encodes four. The register shift delays helix formation, causing overmodification. Differential scanning calorimetry yielded a decrease in Tm of 2 °C for helices with one mutant chain and a 6 °C decrease in helices with two mutant chains. An in vitro binary co-processing assay of N-proteinase cleavage demonstrated that procollagen with the triplet duplication has slower N-propeptide cleavage than in normal controls or procollagen with proα1(I) G832S, G898S, or G997S substitutions, showing that the register shift persists through the entire helix. The register shift disrupts incorporation of mutant collagen into fibrils and matrix. Proband fibrils formed inefficiently in vitro and contained only normal helices and helices with a single mutant chain. Helices with two mutant chains and a significant portion of helices with one mutant chain did not form fibrils. In matrix deposited by proband fibroblasts, mutant chains were abundant in the immaturely cross-linked fraction but constituted a minor fraction of maturely cross-linked chains. The profound effects of shifting the collagen triplet register on chain interactions in the helix and on fibril formation correlate with the severe clinical consequences.

Original languageEnglish
Pages (from-to)10006-10012
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number12
DOIs
StatePublished - Mar 21 2003
Externally publishedYes

Fingerprint

Osteogenesis Imperfecta
Shift registers
Collagen Type I
Extracellular Matrix
Collagen
Mutation
Procollagen
Substitution reactions
Alleles
Differential Scanning Calorimetry
Fibroblasts
Glycine
Differential scanning calorimetry
Exons
Assays
Peptide Hydrolases
Complementary DNA
Processing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Type I collagen triplet duplication mutation in lethal osteogenesis imperfecta shifts register of α chains throughout the helix and disrupts incorporation of mutant helices into fibrils and extracellular matrix. / Cabral, Wayne A.; Mertts, Marianna V.; Makareeva, Elena; Colige, Alain; Tekin, Mustafa; Pandya, Arti; Leikin, Sergey; Marini, Joan C.

In: Journal of Biological Chemistry, Vol. 278, No. 12, 21.03.2003, p. 10006-10012.

Research output: Contribution to journalArticle

Cabral, Wayne A. ; Mertts, Marianna V. ; Makareeva, Elena ; Colige, Alain ; Tekin, Mustafa ; Pandya, Arti ; Leikin, Sergey ; Marini, Joan C. / Type I collagen triplet duplication mutation in lethal osteogenesis imperfecta shifts register of α chains throughout the helix and disrupts incorporation of mutant helices into fibrils and extracellular matrix. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 12. pp. 10006-10012.
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T1 - Type I collagen triplet duplication mutation in lethal osteogenesis imperfecta shifts register of α chains throughout the helix and disrupts incorporation of mutant helices into fibrils and extracellular matrix

AU - Cabral, Wayne A.

AU - Mertts, Marianna V.

AU - Makareeva, Elena

AU - Colige, Alain

AU - Tekin, Mustafa

AU - Pandya, Arti

AU - Leikin, Sergey

AU - Marini, Joan C.

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