Back to Search
Start Over
Cloning and characterization of the heart muscle isoform of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) from crayfish
- Source :
- Syracuse University Experts@Syracuse
- Publication Year :
- 2002
-
Abstract
- SUMMARY This paper describes the cloning and functional characterization of the heart muscle isoform of Sarco/endoplasmic reticulum Ca2+-ATPase(SERCA) from crayfish Procambarus clarkii. The complete crayfish heart SERCA, identified by reverse transcription-polymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends (RACE), consists of 4495 bp with a 3060 bp open reading frame, coding for 1020 amino acids. This isoform differs from the previously identified axial abdominal (tail) muscle SERCA solely in its C-terminal amino acids. The last nine amino acids of the tail muscle isoform are replaced by 27 hydrophobic amino acids in the heart isoform that have the potential to form an additional transmembrane domain. Consistent with other invertebrate studies, Southern blot analysis suggested that the heart and tail muscle isoforms are encoded from the same gene that is equally related to SERCA-1, -2 and -3 of vertebrates. The tissue distributions of these two isoforms have been assessed using isoform-specific probes and northern analysis. A cardiac-specific probe bound only to a 5.8 kb species in heart and had minimal cross-hybridization with 7.6 and 5.8 kb species in eggs and no hybridization with tail muscle. A tail-isoform-specific probe hybridized with a 4.5 kb species in tail muscle and cross-hybridized with a 4.5 kb species in eggs and 8.8 kb in heart muscle. Both isoforms are expressed in eggs suggesting that transcripts are formed early in development and are subsequently broadly expressed in all tissue types. Expression of the cardiac muscle SERCA isoform varied with the stage of moulting. Expression was high in intermoult and decreased in premoult. However, expression was restored rapidly in postmoult (within 2 days) unlike expression of tail muscle SERCA,which remained downregulated for weeks. Differences in contractility between the two muscle types in the postmoult period may explain these expression patterns.
- Subjects :
- Gene isoform
SERCA
DNA, Complementary
Physiology
Molecular Sequence Data
Gene Dosage
Astacoidea
Calcium-Transporting ATPases
Aquatic Science
Endoplasmic Reticulum
Gene Expression Regulation, Enzymologic
Sarcoplasmic Reticulum Calcium-Transporting ATPases
Rapid amplification of cDNA ends
medicine
Animals
Tissue Distribution
Amino Acid Sequence
RNA, Messenger
Cloning, Molecular
Muscle, Skeletal
Molecular Biology
Ecology, Evolution, Behavior and Systematics
chemistry.chemical_classification
Procambarus clarkii
biology
Base Sequence
Endoplasmic reticulum
Myocardium
Cardiac muscle
Gene Expression Regulation, Developmental
biology.organism_classification
Molecular biology
Amino acid
Isoenzymes
Transmembrane domain
Sarcoplasmic Reticulum
medicine.anatomical_structure
chemistry
Insect Science
Animal Science and Zoology
Subjects
Details
- ISSN :
- 00220949
- Volume :
- 205
- Issue :
- Pt 17
- Database :
- OpenAIRE
- Journal :
- The Journal of experimental biology
- Accession number :
- edsair.doi.dedup.....a44eb2e7aea3c0d5eb52e431f8bbd11e