Publications (5)22.96 Total impact
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Article: Kinectin-dependent assembly of translation elongation factor-1 complex on endoplasmic reticulum regulates protein synthesis.
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ABSTRACT: Kinectin is an integral membrane protein with many isoforms primarily found on the endoplasmic reticulum. It has been found to bind kinesin, Rho GTPase, and translation elongation factor-1delta. None of the existing models for the quaternary organization of the elongation factor-1 complex in higher eukaryotes involves kinectin. We have investigated here the assembly of the elongation factor-1 complex onto endoplasmic reticulum via kinectin using in vitro and in vivo assays. We established that the entire elongation factor-1 complex can be anchored to endoplasmic reticulum via kinectin, and the interacting partners are as follows. Kinectin binds EF-1delta, which in turn binds EF-1gamma but not EF-1beta; EF-1gamma binds EF-1delta and EF-1beta but not kinectin. In vivo splice blocking of the kinectin exons 36 and 37 produced kinectin lacking the EF-1delta binding domain, which disrupted the membrane localization of EF-1delta, EF-1gamma, and EF-1beta on endoplasmic reticulum, similar to the disruptions seen with the overexpression of kinectin fragments containing the EF-1delta binding domain. The disruptions of the EF-1delta/kinectin interaction inhibited expression of membrane proteins but enhanced synthesis of cytosolic proteins in vivo. These findings suggest that anchoring the elongation factor-1 complex onto endoplasmic reticulum via EF-1delta/kinectin interaction is important for regulating protein synthesis in eukaryotic cells.Journal of Biological Chemistry 12/2006; 281(44):33621-34. · 4.77 Impact Factor -
Article: Distribution and functions of kinectin isoforms.
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ABSTRACT: Kinectin is an integral transmembrane protein on the endoplasmic reticulum, binding to kinesin, interacting with Rho GTPase and anchoring the translation elongation factor-1 complex. There has been debate on the specific role(s) of kinectin in different species and cell types. Here we identified 15 novel kinectin isoforms in the mouse nervous system, constituting a family of alternatively spliced carboxyl-terminal variants. Isoform expression is subject to cell type- and developmental stage-specific regulation. We raised specific antibodies to the kinectin variants to characterise their differential intracellular localisation and discovered that certain kinectin isoforms are found in axons where kinectin was previously believed to be absent. We also demonstrated in vivo by overexpression and RNA interference assay that kinectin is selectively involved in the transport of specific types of organelles. A 160 kDa kinectin species is mainly concentrated in the endoplasmic reticulum, anchored via its transmembrane domain and is essential for endoplasmic reticulum membrane extension. A 120 kDa kinectin species is specifically associated with mitochondria, and its interaction with kinesin was found to influence mitochondrial dynamics. These findings contribute to a more unified view of kinectin function. They suggest that different cellular processes use specific kinectin isoforms to mediate intracellular motility and targeting by transient interaction with different motor proteins or other binding partners.Journal of Cell Science 10/2004; 117(Pt 19):4537-49. · 6.11 Impact Factor -
Article: Cloning of rat telomerase catalytic subunit functional domains, reconstitution of telomerase activity and enzymatic profile of pig and chicken tissues.
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ABSTRACT: Telomerase is a ribonucleoprotein polymerase which adds TTAGGG repeats to telomeric ends. Recent studies reported the reverse transcription enzyme activity mostly from the catalytic subunit (TERT) of the enzyme complex. Both human telomerase catalytic subunit (hTERT) and mouse telomerase catalytic subunit (mTERT) had been previously cloned but not rat telomerase catalytic subunit rTERT. In this study, the rTERT functional domains were cloned and was found that its function resemble to mouse and human telomerase. In addition, chicken and pig telomerase activity profile were studied and its enzyme activity is related to its proliferation capability of individual tissues. However, its catalytic subunit does not like mouse, rat and human cases that the telomerase activity could not reconstituted by the in-vitro transfection of mTERT and hTERT cloned vectors. Here we demonstrated that rTERT is similar to mTERT and hTERT but not pig and chicken telomerase. Further studies are needed to verify the malignancy characteristics because nowadays artificial organs/tissues from these animals are used for the transplantation to human body.Life Sciences 11/2003; 73(21):2749-60. · 2.53 Impact Factor -
Article: Kinectin anchors the translation elongation factor-1 delta to the endoplasmic reticulum.
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ABSTRACT: Kinectin has been proposed to be a membrane anchor for kinesin on intracellular organelles. A kinectin isoform that lacks a major portion of the kinesin-binding domain does not bind kinesin but interacts with another resident of the endoplasmic reticulum, the translation elongation factor-1 delta (EF-1 delta). This was shown by yeast two-hybrid analysis and a number of in vitro and in vivo assays. EF-1 delta provides the guanine nucleotide exchange activities on EF-1 alpha during elongation step of protein synthesis. The minimal EF-1 delta-binding domain on kinectin resides within a conserved region present in all the kinectin isoforms. Overexpression of the kinectin fragments in vivo disrupted the intracellular localization of EF-1 delta proteins. This report provides evidence of an alternative kinectin function as the membrane anchor for EF-1 delta on the endoplasmic reticulum and provides clues to the EF-1 complex assembly and anchorage on the endoplasmic reticulum.Journal of Biological Chemistry 09/2003; 278(34):32115-23. · 4.77 Impact Factor -
Article: Kinectin-Kinesin Binding Domains and Their Effects on Organelle Motility
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ABSTRACT: Intracellular organelle motility involves motor proteins that move along microtubules or actin filaments. One of these motor proteins, kinesin, was proposed to bind to kinectin on membrane organelles during movement. Whether kinectin is the kinesin receptor on organelles with a role in organelle motility has been controversial. We have characterized the sites of interaction between human kinectin and conventional kinesin using in vivo and in vitro assays. The kinectin-binding domain on the kinesin tail partially overlaps its head-binding domain and the myosin-Va binding domain. The kinesin-binding domain on kinectin resides near the COOH terminus and enhances the microtubule-stimulated kinesin-ATPase activity, and the overexpression of the kinectin-kinesin binding domains inhibited kinesin-dependent organelle motility in vivo. These data, when combined with other studies, suggest a role for kinectin in organelle motility.Journal of Biological Chemistry 10/2000; 275(42):32854-32860. · 4.77 Impact Factor
Top co-authors
Top Journals
Institutions
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2003–2006
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National University of Singapore
- Department of Physiology
Singapore, Singapore
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2004
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University of Cyprus
Nicosia, Nicosia District, Cyprus
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