Cloning and functional characterization of an uncoupling protein homolog in hummingbirds

Claudia R. Vianna, Thilo Hagen, Chen Yu Zhang, Eric Bachman, Olivier Boss, B. Gereben, Anselmo S. Moriscot, Bradford B. Lowell, José Eduardo P W Bicudo, Antonio C. Bianco

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room's temperature to 12-14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.

Original languageEnglish
Pages (from-to)137-145
Number of pages9
JournalPhysiological Genomics
Volume2001
Issue number5
Publication statusPublished - Jun 2001

Fingerprint

Organism Cloning
Torpor
Pectoralis Muscles
Rewarming
Complementary DNA
Open Reading Frames
Mitochondrial Uncoupling Proteins
Yeasts
Swallows
Messenger RNA
Proteins
Mitochondrial Membrane Potential
Iodides
Deglutition
Northern Blotting
Birds
Skeletal Muscle
Coloring Agents
Nucleotides
Western Blotting

Keywords

  • Birds
  • Brown adipose tissue
  • Nonshivering thermogenesis
  • Torpor

ASJC Scopus subject areas

  • Genetics
  • Physiology

Cite this

Vianna, C. R., Hagen, T., Zhang, C. Y., Bachman, E., Boss, O., Gereben, B., ... Bianco, A. C. (2001). Cloning and functional characterization of an uncoupling protein homolog in hummingbirds. Physiological Genomics, 2001(5), 137-145.

Cloning and functional characterization of an uncoupling protein homolog in hummingbirds. / Vianna, Claudia R.; Hagen, Thilo; Zhang, Chen Yu; Bachman, Eric; Boss, Olivier; Gereben, B.; Moriscot, Anselmo S.; Lowell, Bradford B.; Bicudo, José Eduardo P W; Bianco, Antonio C.

In: Physiological Genomics, Vol. 2001, No. 5, 06.2001, p. 137-145.

Research output: Contribution to journalArticle

Vianna, CR, Hagen, T, Zhang, CY, Bachman, E, Boss, O, Gereben, B, Moriscot, AS, Lowell, BB, Bicudo, JEPW & Bianco, AC 2001, 'Cloning and functional characterization of an uncoupling protein homolog in hummingbirds', Physiological Genomics, vol. 2001, no. 5, pp. 137-145.
Vianna CR, Hagen T, Zhang CY, Bachman E, Boss O, Gereben B et al. Cloning and functional characterization of an uncoupling protein homolog in hummingbirds. Physiological Genomics. 2001 Jun;2001(5):137-145.
Vianna, Claudia R. ; Hagen, Thilo ; Zhang, Chen Yu ; Bachman, Eric ; Boss, Olivier ; Gereben, B. ; Moriscot, Anselmo S. ; Lowell, Bradford B. ; Bicudo, José Eduardo P W ; Bianco, Antonio C. / Cloning and functional characterization of an uncoupling protein homolog in hummingbirds. In: Physiological Genomics. 2001 ; Vol. 2001, No. 5. pp. 137-145.
@article{0d1760a4e11f4f0ea655a883efa7a27a,
title = "Cloning and functional characterization of an uncoupling protein homolog in hummingbirds",
abstract = "The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55{\%} identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room's temperature to 12-14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.",
keywords = "Birds, Brown adipose tissue, Nonshivering thermogenesis, Torpor",
author = "Vianna, {Claudia R.} and Thilo Hagen and Zhang, {Chen Yu} and Eric Bachman and Olivier Boss and B. Gereben and Moriscot, {Anselmo S.} and Lowell, {Bradford B.} and Bicudo, {Jos{\'e} Eduardo P W} and Bianco, {Antonio C.}",
year = "2001",
month = "6",
language = "English",
volume = "2001",
pages = "137--145",
journal = "Physiological Genomics",
issn = "1531-2267",
publisher = "American Physiological Society",
number = "5",

}

TY - JOUR

T1 - Cloning and functional characterization of an uncoupling protein homolog in hummingbirds

AU - Vianna, Claudia R.

AU - Hagen, Thilo

AU - Zhang, Chen Yu

AU - Bachman, Eric

AU - Boss, Olivier

AU - Gereben, B.

AU - Moriscot, Anselmo S.

AU - Lowell, Bradford B.

AU - Bicudo, José Eduardo P W

AU - Bianco, Antonio C.

PY - 2001/6

Y1 - 2001/6

N2 - The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room's temperature to 12-14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.

AB - The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room's temperature to 12-14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.

KW - Birds

KW - Brown adipose tissue

KW - Nonshivering thermogenesis

KW - Torpor

UR - http://www.scopus.com/inward/record.url?scp=0348203045&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0348203045&partnerID=8YFLogxK

M3 - Article

VL - 2001

SP - 137

EP - 145

JO - Physiological Genomics

JF - Physiological Genomics

SN - 1531-2267

IS - 5

ER -