Identification and characterization of cvHsp. A novel human small stress protein selectively expressed in cardiovascular and insulin-sensitive tissues.
Starting with computational tools that search for tissue-selective expression of assembled expressed sequenced tags, we have identified by focusing on heart libraries a novel small stress protein of 170 amino acids that we named cvHsp. cvHsp was found as being computationally selectively and highly (0.3% of the total RNA) expressed in human heart. The cvHsp gene mapped to 1p36.23-p34.3 between markers D1S434 and D1S507. The expression of cvHsp was analyzed with RNA dot, Northern blots, or reverse transcription-polymerase chain reaction: expression was high in heart, medium in skeletal muscle, and low in aorta or adipose tissues. In the heart of rat models of cardiac pathologies, cvHsp mRNA expression was either unchanged (spontaneous hypertension), up-regulated (right ventricular hypertrophy induced by monocrotaline treatment), or down-regulated (left ventricular hypertrophy following aortic banding). In obese Zucker rats, cvHsp mRNA was increased in skeletal muscle, brown, and white adipose tissues but remained unchanged in the heart. Western blot analysis using antipeptide polyclonal antibodies revealed two specific bands at 23 and 25 kDa for cvHsp in human heart. cvHsp interacted in both yeast two-hybrid and immunoprecipitation experiments with alpha-filamin or actin-binding protein 280. Within cvHsp, amino acid residues 56-119 were shown to be important for its specific interaction with the C-terminal tail of alpha-filamin.
Krief
S
S
SmithKline Beecham Laboratoires Pharmaceutiques, 4 rue du Chesnay-Beauregard, BP 58, 35762 Saint-Grégoire, France. Stephane_Krief@sbphrd.com
Faivre
J F
JF
Robert
P
P
Le Douarin
B
B
Brument-Larignon
N
N
Lefrère
I
I
Bouzyk
M M
MM
Anderson
K M
KM
Greller
L D
LD
Tobin
F L
FL
Souchet
M
M
Bril
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002319:Cardiovascular System / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D051381:Rats; D016415:Sequence Alignment
D018076:DNA, Complementary; D006360:Heat-Shock Proteins; D007328:Insulin