Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo.

Agah, Ramtin; Frenkel, Peter A.; French, Brent A.; Michael, L H; Overbeek, Paul A.; Schneider, Michel

doi:10.1172/jci119509

Cited by 547 publications

(527 citation statements)

References 49 publications

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“…To generate the heart-specific Rce1 knockout mice, we bred Rce1 flx/flx mice harboring a Cre transgene under the control of the ␣Myhc promoter (22,30). As expected, Southern blots of genomic DNA from the hearts of Rce1 flx/flx ␣Myhc-Cre ϩ/o mice revealed incomplete recombination ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…As expected from earlier studies (6), mice with a single copy of the knockout allele (Rce1 ⌬/ϩ ) were phenotypically normal and free of pathology over 18 months of observation. Mice hemizygous for an ␣-myosin heavy chain-Cre transgene (␣Myhc-Cre ϩ/o ) (22) were obtained from Dr. Michael Schneider (Baylor College of Medicine, Houston, TX).…”

Section: Methodsmentioning

confidence: 99%

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On the Physiological Importance of Endoproteolysis of CAAX Proteins

Bergö

Lieu

Gavino

et al. 2004

Journal of Biological Chemistry

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Proteins terminating with a CAAX motif, such as the Ras proteins and the nuclear lamins, undergo posttranslational modification of a C-terminal cysteine with an isoprenyl lipid via a process called protein prenylation. After prenylation, the last three residues of CAAX proteins are clipped off by Rce1, an integral membrane endoprotease of the endoplasmic reticulum. Prenylation is crucial to the function of many CAAX proteins, but the physiologic significance of endoproteolytic processing has remained obscure. To address this issue, we used Cre/loxP recombination techniques to create mice lacking Rce1 in the heart, an organ where Rce1 is expressed at particularly high levels. The hearts from heart-specific Rce1 knockout mice manifested reduced levels of both the Rce1 mRNA and CAAX endoprotease activity, and the hearts manifested an accumulation of CAAX protein substrates. The heart-specific Rce1 knockout mice initially appeared healthy but died starting at 3-5 months of age. By 10 months of age, ϳ70% of the mice had died. Pathological studies revealed that the heartspecific Rce1 knockout mice had a dilated cardiomyopathy. By contrast, liver-specific Rce1 knockout mice appeared healthy, had normal transaminase levels, and had normal liver histology. These studies indicate that the endoproteolytic processing of CAAX proteins is essential for cardiac function but is less important for the liver.Proteins terminating with a CAAX 1 motif, such as the nuclear lamins and the Ras and Rho proteins, undergo three sequential post-translational processing steps (1-4). First, the C-terminal cysteine (i.e. the C of the CAAX motif) is "lipidated" with a 15-carbon farnesyl or a 20-carbon geranylgeranyl lipid. This processing step is carried out by cytosolic prenyltransferases (protein farnesyltransferase and protein geranylgeranyltransferase type I). Second, a prenylprotein-specific endoprotease of the endoplasmic reticulum, Ras-converting enzyme 1 (Rce1), clips off the last three amino acids from the protein (i.e. the -AAX) (5-7). Third, the newly exposed C-terminal isoprenylcysteine is methylesterified by isoprenylcysteine carboxyl methyltransferase (Icmt), a prenylprotein-specific, S-adenosylmethionine-dependent methyltransferase of the endoplasmic reticulum (8 -11).Prenylation of CAAX proteins is vitally important to eukaryotic cells (1-4). Yeast lacking the shared ␣-chain of farnesyltransferase and geranylgeranyltransferase type I are not viable (12). In mammalian cells, prenylation of CAAX proteins is absolutely required for their proper targeting to membrane surfaces and for proper protein function (1-4). The importance of protein farnesylation is clearly indicated by the fact that mice lacking farnesyltransferase die early during embryonic development (before embryonic day 6.5). 2 The geranylgeranylation of CAAX proteins is also critical for normal cellular function. Drugs that inhibit geranylgeranylation of CAAX proteins have pronounced effects on cell growth and can trigger apoptosis (13-15). Further underscoring the im...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

On the Physiological Importance of Endoproteolysis of CAAX Proteins

Bergö

Lieu

Gavino

et al. 2004

Journal of Biological Chemistry

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“…Transgenic mice expressing Cre recombinase under the control of the α-myosin heavy chain promoter (αMyHC-Cre) were created as described previously [11]. Homozygous β-catenin loxP-floxed (β-catenin fl/fl ) mice were crossed with αMyHC-Cre mice to generate heterozygous β-catenin loxPfloxed (β-catenin fl/wt ) negative for αMyHC-Cre or heterozygous β-catenin deleted (β-catenin del/wt ) positive for αMyHC-Cre.…”

Section: Animalsmentioning

confidence: 99%

“…DNA was isolated from tail or toe biopsy of neonates following proteinase K digestion as described [10]. αMyHC-Cre transgene, loxP-floxed and wild type β-catenin genes were amplified by PCR as described previously [10,11]. A pilot study confirmed that recombination of β-catenin gene only occurred in atrial and ventricular tissues.…”

Section: Animalsmentioning

confidence: 99%

Cardiac-specific haploinsufficiency of β-catenin attenuates cardiac hypertrophy but enhances fetal gene expression in response to aortic constriction

Zhou

et al. 2007

Journal of Molecular and Cellular Cardiology

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In addition to its role in cell adhesion, β-catenin is an important signaling molecule in the Wnt/ Wingless signaling pathway. Recent studies have indicated that β-catenin is stabilized by hypertrophic stimuli and may regulate cardiac hypertrophic responses. To explore the role and requirement of β-catenin in cardiac development and hypertrophy, we deleted the β-catenin gene specifically in cardiac myocytes by crossing loxP-floxed β-catenin mice with transgenic mice expressing a Cre recombinase under the control of the α-myosin heavy chain promoter. No homozygous β-catenin deleted mice were born alive and died before embryonic day 14.5, indicating significant and irreplaceable roles of β-catenin in embryonic heart development. Heterozygous β-catenin deleted mice, however, demonstrated no structural and functional abnormality. The response of heterozygous β-catenin deleted mice to transverse aortic constriction, however, was significantly attenuated with decreased heart weight and heart weight/body weight ratio compared to controls with intact β-catenin genes. Hemodynamic analysis revealed that there was no difference in cardiac function between wild type and heterozygous β-catenin deleted mice. On the other hand, the expression of fetal genes, β-myosin heavy chain, atrial and brain natriuretic peptides was significantly higher in heterozygous β-catenin deleted mice when compared to wild type β-catenin mice. These results suggest that the cytoplasmic level of β-catenin modulates hypertrophic response and fetal gene reprogramming after pressure overload.

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“…As an initial step to help distinguish the lineage responsible for the Cx43-null phenotype, we have previously used a targeted approach to delete Cx43 expression specifically in cardiomyocytes with two separate cardiac-specific Cre recombinase mouse lines, MLC-2v-Cre (Chen et al, 1998) and ␣-MHC-Cre (Agah et al, 1997). Both MLC-2v-Cre and ␣-MHC-Cre are expressed in the heart prior to E9 (Chen et al, 1998;de Lange et al, 2004).…”

Section: Research Articlementioning

confidence: 99%

Distinct cardiac malformations caused by absence of connexin 43 in the neural crest and in the non-crest neural tube

Liu

Schneider

et al. 2006

Development

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Connexin 43 (Cx43) is expressed in the embryonic heart, cardiac neural crest (CNC) and neural tube, and germline knockout (KO) of Cx43 results in aberrant cardiac outflow tract (OFT) formation and abnormal coronary deployment. Prior studies suggest a vital role for CNC expression of Cx43 in heart development. Surprisingly, we found that conditional knockout (CKO) of Cx43 in the dorsal neural tube and CNC mediated by Wnt1-Cre failed to recapitulate the Cx43-null OFT phenotype, although coronary vasculature was abnormal in this mutant line. A broader CKO mediated by P3pro (Pax3)-Cre, involving both ventral and dorsal aspects of the thoracic neural tube and CNC, resulted in infundibular bulging and coronary anomalies similar to those seen in germline Cx43-null hearts. P3pro-Cre-mediated loss of Cx43 in the neural tube was characterized by a late phase of cellular delamination from the dorsal and lateral neural tube, a markedly increased abundance of neuroepithelium-derived cells outside of the neural tube and an excess of such cells infiltrating the heart and infundibulum. Thus, expression of Cx43 in the CNC is crucial for normal coronary deployment, but Cx43 is not required in the CNC for normal OFT morphogenesis. Rather, this study suggests a novel function for Cx43 in which Cx43 acts through non-crest neuroepithelial cells to suppress cellular delamination from the neural tube and thereby preserve normal OFT development.

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Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo.

Cited by 547 publications

References 49 publications

On the Physiological Importance of Endoproteolysis of CAAX Proteins

On the Physiological Importance of Endoproteolysis of CAAX Proteins

Cardiac-specific haploinsufficiency of β-catenin attenuates cardiac hypertrophy but enhances fetal gene expression in response to aortic constriction

Distinct cardiac malformations caused by absence of connexin 43 in the neural crest and in the non-crest neural tube

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