Molecular Cloning and Characterization of SmLIM, a Developmentally Regulated LIM Protein Preferentially Expressed in Aortic Smooth Muscle Cells

Jain, Mukesh K.; Fujita, Kenji; Hsieh, Chung Ming; Endege, Wilson O.; Sibinga, Nicholas E.S.; Yet, Shaw‐Fang; Kashiki, Saori; Lee, Wen Sen; Perrella, Mark A.; Haber, Edgar; Lee, Mu En

doi:10.1074/jbc.271.17.10194

Cited by 89 publications

(102 citation statements)

References 42 publications

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“…RNA Preparation, Northern Blot Analysis, and in Situ Hybridization-Total RNA was prepared from mouse organ tissue and cultured cells as described (33). Northern blot analysis was performed as described (33).…”

Section: Methodsmentioning

confidence: 99%

“…Northern blot analysis was performed as described (33). In situ hybridization was performed as described (33).…”

Section: Methodsmentioning

confidence: 99%

“…Northern blot analysis was performed as described (33). In situ hybridization was performed as described (33). The mCHF1 riboprobe was derived from a 0.5-kb EcoRI to Acc65I fragment encoding the bHLH domain cloned into pBluescript II SK.…”

Section: Methodsmentioning

confidence: 99%

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Cardiovascular Basic Helix Loop Helix Factor 1, a Novel Transcriptional Repressor Expressed Preferentially in the Developing and Adult Cardiovascular System

Chin

Maemura

Fukumoto

et al. 2000

Journal of Biological Chemistry

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143

132

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We have cloned a cardiovascular-restricted basic helixloop-helix factor that interacts with arylhydrocarbon receptor nuclear translocator (ARNT) in a yeast two-hybrid screen. Cardiovascular helix-loop-helix factor 1 (CHF1) is distantly related to the hairy family of transcriptional repressors. We analyzed its expression pattern during mouse embryo development. At day 8.5, the expression of CHF1 is first detected in the primitive ventricle of the primordial heart tube and persists throughout gestation. In rat hearts, this expression is down-regulated after birth, concurrent with terminal differentiation of cardiomyocytes. In the developing vasculature, CHF1 first appears in the dorsal aorta at day 9.0, which precedes the reported expression of smooth muscle cell markers, and persists into adulthood. In an in vitro system of smooth muscle cell differentiation, CHF1 mRNA was barely detectable in undifferentiated cells but was induced highly in differentiated smooth muscle cells. To determine whether CHF1 might affect the function of ARNT, we performed transfection studies. Co-transfection of CHF1 inhibited ARNT/EPAS1-dependent transcription by 85%, and this inhibition is dose-dependent. In electrophoretic mobility studies, CHF1 inhibited the binding of the ARNT/EPAS1 heterodimer to its target site. Our data suggest that CHF1 functions as a transcriptional repressor and may play an important role in cardiovascular development.

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Section: Methodsmentioning

confidence: 99%

“…Northern blot analysis was performed as described (33). In situ hybridization was performed as described (33).…”

Section: Methodsmentioning

confidence: 99%

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Cardiovascular Basic Helix Loop Helix Factor 1, a Novel Transcriptional Repressor Expressed Preferentially in the Developing and Adult Cardiovascular System

Chin

Maemura

Fukumoto

et al. 2000

Journal of Biological Chemistry

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143

132

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“…These proteins are CRP1 (29,48), CRP2/SmLIM (38,49), and CRP3/ MLP (39). There is growing evidence that CRPs mediate protein-protein interaction by their two LIM domains (50,51).…”

Section: Figmentioning

confidence: 99%

Cysteine-rich Protein 2, a Novel Substrate for cGMP Kinase I in Enteric Neurons and Intestinal Smooth Muscle

Huber¹,

Neuhuber²,

Klugbauer³

et al. 2000

Journal of Biological Chemistry

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Nitric oxide/cGMP/cGMP kinase I (cGKI) signaling causes relaxation of intestinal smooth muscle. In the gastrointestinal tract substrates of cGKI have not been identified yet. In the present study a protein interacting with cGKI␤ has been isolated from a rat intestinal cDNA library using the yeast two-hybrid system. The protein was identified as cysteine-rich protein 2 (CRP2), recently cloned from rat brain (Okano, I., Yamamoto, T., Kaji, A., Kimura, T., Mizuno, K., and Nakamura, T. (1993) FEBS Lett. 333, 51-55). Recombinant CRP2 is specifically phosphorylated by cGKs but not by cAMP kinase in vitro. Co-transfection of CRP2 and cGKI␤ into COS cells confirmed the phosphorylation of CRP2 in vivo. Cyclic GMP kinase I phosphorylated CRP2 at Ser-104, because the mutation to Ala completely prevented the in vivo phosphorylation. Immunohistochemical analysis using confocal laser scan microscopy showed a co-localization of CRP2 and cGKI in the inner part of the circular muscle layer, in the muscularis mucosae, and in specific neurons of the myenteric and submucosal plexus. The co-localization together with the specific phosphorylation of CRP2 by cGKI in vitro and in vivo suggests that CRP2 is a novel substrate of cGKI in neurons and smooth muscle of the small intestine.Nitric oxide, a candidate of nonadrenergic, noncholinergic neurotransmission of the enteric nervous system (1, 2) has a variety of physiological functions in the gut (3, 4). The inhibitory effect of NO on smooth muscle cells, either of the gut wall (5) or intestinal blood vessels (6), regulates gut motility and blood flow. Further, NO modulates neurosecretion from enteric nerve terminals (7). A main signaling pathway of NO in the gut is the activation of soluble guanylyl cyclase (8 -10), which results in an increase of intracellular cGMP. Cyclic GMP analogues elicit NO-like effects on smooth muscle (11) and isolated nerve terminals (7) causing smooth muscle relaxation and release of vasoactive intestinal polypeptide, respectively. A major target of cGMP in smooth muscle is the cGMP-dependent protein kinase I (cGKI) 1 (12-14). The two existing isoforms of cGKI, cGKI␣ and cGKI␤, show a tissue-specific distribution (15), with cGKI␤ being the predominant isoform in the small intestine (5). (22) and on the stimulation of the large conductance K Ca channel (BK) (23,24). Furthermore, the relaxation of smooth muscle by NO/cGMP is also associated with an increase in the extent of phosphorylation of small proteins like heat shock related proteins that appear to be regulatory components of the actin based cytoskeleton or that seem to interact with intermediate filaments (25). A further substrate of cGKI involved in actin filament assembling and cell motility is the vasodilator-stimulated phosphoprotein associated with focal adhesions and areas of dynamic membrane activity (26). These proteins may be involved in relaxation and contractions of vascular smooth muscle by regulating cytoskeletal organization and cell shape. In contrast to vascular smooth muscle, little is...

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“…Studies in both chick and rat have revealed that CRP2 (also referred to as SmLIM) is present at highest levels in arterial samples (Jain et al 1996;Louis et al 1997), and subsequent work has focused on a potential role for CRP2 in the vascular system (Jain et al 1998). Here we describe the pattern of csrp2 expression in the developing and adult mouse.…”

mentioning

confidence: 99%

Expression of the Gene Encoding the LIM Protein CRP2: A Developmental Profile

Henderson

Brown

Richardson

et al. 2002

J Histochem Cytochem.

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(JAR) S U M M A R Y Members of the cysteine-rich protein (CRP) family are evolutionarily conserved L in-11, I sl-1, M ac-3 (LIM) domain proteins that have been implicated in cell differentiation. Here we describe the expression pattern of the CRP family member CRP2 in mouse. Unlike other CRP family members, which are expressed primarily in muscle, CRP2 is more broadly expressed. In addition to expression in vascular smooth muscle cells, we also detect CRP2-specific transcripts in mesenchymal derivatives and several epithelia.

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Molecular Cloning and Characterization of SmLIM, a Developmentally Regulated LIM Protein Preferentially Expressed in Aortic Smooth Muscle Cells

Cited by 89 publications

References 42 publications

Cardiovascular Basic Helix Loop Helix Factor 1, a Novel Transcriptional Repressor Expressed Preferentially in the Developing and Adult Cardiovascular System

Cardiovascular Basic Helix Loop Helix Factor 1, a Novel Transcriptional Repressor Expressed Preferentially in the Developing and Adult Cardiovascular System

Cysteine-rich Protein 2, a Novel Substrate for cGMP Kinase I in Enteric Neurons and Intestinal Smooth Muscle

Expression of the Gene Encoding the LIM Protein CRP2: A Developmental Profile

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