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United States Department of Agriculture

Agricultural Research Service

Title: Increased Type I Collagen Content and DNA Binding Activity of a Single-Stranded Cytosine-Rich Sequence in the Nuclear Extract of the Copper-Deficient Rat Heart

Authors
item Zeng, Huawei
item Saari, Jack

Submitted to: Journal of Federation of American Societies for Experimental Biology
Publication Type: Abstract Only
Publication Acceptance Date: December 1, 2003
Publication Date: March 23, 2004
Citation: Zeng, H., Saari, J.T. 2004. Increased type I collagen content and DNA binding activity of a single-stranded cytosine-rich sequence in the nuclear extract of the copper-deficient rat heart [abstract]. The Federation of American Societies for Experimental Biology Journal. 18:A914.

Interpretive Summary: Dietary Cu-deficiency not only causes a hypertrophic cardiomyopathy but also increases cancer risk in rodent models. However, a possible alteration in gene expression has not been fully examined. The present study was undertaken to determine the effect of Cu-deficiency on protein profiles in rat heart tissue. Male Sprague-Dawley rats were fed diets that were either copper adequate (6.0 ug copper/g diet n=6) or copper deficient (0.3 ug copper/g diet n=6) for 5 wk. Nuclear extracts from heart tissue of Cu-deficient, but not Cu-adequate rats showed a 132 kDa protein complex by SDS-PAGE analysis. This complex stained pink with Coomassie Blue, suggesting the presence of collagens or other proline-rich proteins. Dot immunoblotting demonstrated that total type I collagen was increased by 110% in the nuclear extract from Cu-deficient, relative to Cu-adequate animals. LCMS analysis indicated that the 132 kDa protein complex contained a collagen alpha (I) chain precursor as well as a leucine-rich protein 130 in the nuclear extract from Cu-deficient, but not Cu-adequate rats. A gel shift assay showed that nuclear extract from Cu-deficient rats bound to a single-stranded cytosine-rich sequence with higher affinity than the extract of Cu-adequate rats The data demonstrate that Cu adequacy is necessary for the synthesis and deposition of type I collagen and appears to be necessary for maintaining genome DNA integrity.

Technical Abstract: Dietary Cu-deficiency not only causes a hypertrophic cardiomyopathy but also increases cancer risk in rodent models. However, a possible alteration in gene expression has not been fully examined. The present study was undertaken to determine the effect of Cu-deficiency on protein profiles in rat heart tissue. Male Sprague-Dawley rats were fed diets that were either copper adequate (6.0 ug copper/g diet n=6) or copper deficient (0.3 ug copper/g diet n=6) for 5 wk. Nuclear extracts from heart tissue of Cu-deficient, but not Cu-adequate rats showed a 132 kDa protein complex by SDS-PAGE analysis. This complex stained pink with Coomassie Blue, suggesting the presence of collagens or other proline-rich proteins. Dot immunoblotting demonstrated that total type I collagen was increased by 110% in the nuclear extract from Cu-deficient, relative to Cu-adequate animals. LCMS analysis indicated that the 132 kDa protein complex contained a collagen alpha (I) chain precursor as well as a leucine-rich protein 130 in the nuclear extract from Cu-deficient, but not Cu-adequate rats. A gel shift assay showed that nuclear extract from Cu-deficient rats bound to a single-stranded cytosine-rich sequence with higher affinity than the extract of Cu-adequate rats The data demonstrate that Cu adequacy is necessary for the synthesis and deposition of type I collagen and appears to be necessary for maintaining genome DNA integrity.

Last Modified: 7/25/2014
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