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onsdag 6 november 2019

KLHL1(13q21.33) , MRP2. Oligodendrosyytille tärkeä Mayvenin sukuinen MRP2 = Kelch proteiini 1)

http://www.jbc.org/content/282/16/12319.long
 Tutkijat luonnehtivat uuden Kelch proteiinin joka on KLHL1 kaltainen ja Mayvenin sukuinen MPR2.  Se osoittautuu tärkeäksi merkitsijäksi oligodenrosyyttien elämässä.

Process Elongation of Oligodendrocytes Is Promoted by the Kelch-related Protein MRP2/KLHL1*
Shuxian Jiang1, Seyha Seng1, Hava Karsenty Avraham, Yigong Fu and Shalom Avraham2
  1. Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115
  1. 2 To whom correspondence should be addressed: 4 Blackfan Circle, Boston, MA 02115. Tel.: 617-667-0063; Fax: 617-975-6373 or 617-975-5240; E-mail: savraham{at}bidmc.harvard.edu.
Abstract
Oligodendrocytes (OLGs) are generated by progenitor cells that are committed to differentiating into myelin-forming cells of the central nervous system. Rearrangement of the cytoskeleton leading to the extension of cellular processes is essential for the myelination of axons by OLGs. Here, we have characterized a new member of the Kelch-related protein family termed MRP2 (for Mayven-related protein 2) that is specifically expressed in brain. MRP2/KLHL1 is expressed in oligodendrocyte precursors and mature OLGs, and its expression is up-regulated during OLG differentiation. MRP2/KLHL1 expression was abundant during the specific stages of oligodendrocyte development, as identified by A2B5-, O4-, and O1-specific oligodendrocyte markers. MRP2/KLHL1 was localized in the cytoplasm and along the cell processes. Moreover, a direct endogenous association of MRP2/KLHL1 with actin was observed, which was significantly increased in differentiated OLGs compared with undifferentiated OLGs. Overexpression of MRP2/KLHL1 resulted in a significant increase in the process extension of rat OLGs, whereas MRP2/KLHL1 antisense reduced the process length of primary rat OLGs. Furthermore, murine OLGs isolated from MRP2/KLHL1 transgenic mice showed a significant increase in the process extension of OLGs compared with control wild-type murine OLGs. These studies provide insights into the role of MRP2/KLHL1, through its interaction with actin, in the process elongation of OLGs.

Oligodendrocytes (OLGs)3 are a major cell type in the central nervous system. Development of these cells is necessary for normal functioning of the brain, and injury to them is involved in the pathogenesis of important neurological disorders including cerebral palsy, multiple sclerosis, and periventricular leukomalacia (1, 2). OLGs represent the myelin-forming cells of the central nervous system. They produce numerous membranous processes, which spirally enwrap neuronal axons, forming multilamellar myelin sheaths (3, 4). OLGs are metabolically the most active cells in the brain (5). Before OLGs can remyelinate, they must first be able to extend their processes, and contact the demyelinated axons. However, the molecules involved in the mechanisms of OLG process extension are poorly defined.
A new and unique family of actin-binding proteins with sequences and domains homologous with the Drosophila “Kelch” protein has emerged (6). Kelch protein is believed to be important for the maintenance of the ordered cytoskeleton (7, 8). The Kelch protein has two structural domains that are also found in other molecules. The first domain, which consists of about 115 amino acids, has been named the BTB (Bric-a-brac, Tramtrack, Broad-complex) domain (9) or POZ (Poxvirus zinc finger) domain (10). The second domain, composed of about 50 amino acids repeated in tandem, has been called the “Kelch repeats.” The BTB/POZ domain has been proposed to function as a protein-protein interaction interface, which organizes higher order structures involved in chromatin folding or cytoskeleton organization (11). 


The Kelch-related proteins are a superfamily of proteins conserved in a wide range of organisms, from viruses to mammals. At least 60 Kelch-related proteins have been identified, but their physiological and biochemical functions remain largely uncharacterized (12, 13). The Drosophila Kelch proteins colocalize with actin filaments in a structure called the ring canal, which bridges 15 nurse cells and the oocyte. Drosophila Kelch protein plays an important role in maintaining actin organization during the development of ring canals (6, 8).

The Kelch-related proteins have diverse functions in cell morphology, cell organization, and gene expression, and function in multiprotein complexes through contact sites in their β-propeller domains (14). Recently, a new member of the BTB/Kelch repeat family, gigaxonin (GAN, KLHL16), was reported to be a pathological target for neurodegenerative disorders in which alterations were found to contain multiple mutations in the Kelch repeats in the neurofilament network (15).
We have previously identified and characterized two actinbinding proteins, termed NRP/B/ENC-1 (KLHL37, PIG10 TP5310 ,5q13.3) (16-18) and Mayven (KHLH24q32.3)  (19), predominantly expressed in brain. Mayven is an actin-binding protein that is co-localized with actin filaments in stress fibers and in the patchy cortical actin-rich regions of the cell margins and processes, including the process tips in primary neurons and U373-MG astrocytoma/glioblastoma cells (19). During our study of proteins that are related to Mayven, we identified and cloned a novel gene, which we termed: MRP2 (Mayven-related protein 2) that was found to be identical to KLHL1 (20, 21). In this study, we have investigated the expression of MRP2/KLHL1 in OLGs and its possible role in the dynamics of cytoskeletal rearrangement, leading to the elongation of OLG processes.

Oligodendrocytes (OLGs)3 are a major cell type in the central nervous system. Development of these cells is necessary for normal functioning of the brain, and injury to them is involved in the pathogenesis of important neurological disorders including cerebral palsy, multiple sclerosis, and periventricular leukomalacia (1, 2). OLGs represent the myelin-forming cells of the central nervous system. They produce numerous membranous processes, which spirally enwrap neuronal axons, forming multilamellar myelin sheaths (3, 4). OLGs are metabolically the most active cells in the brain (5). Before OLGs can remyelinate, they must first be able to extend their processes, and contact the demyelinated axons. However, the molecules involved in the mechanisms of OLG process extension are poorly defined.
A new and unique family of actin-binding proteins with sequences and domains homologous with the Drosophila “Kelch” protein has emerged (6). Kelch protein is believed to be important for the maintenance of the ordered cytoskeleton (7, 8). The Kelch protein has two structural domains that are also found in other molecules. The first domain, which consists of about 115 amino acids, has been named the BTB (Bric-a-brac, Tramtrack, Broad-complex) domain (9) or POZ (Poxvirus zinc finger) domain (10). The second domain, composed of about 50 amino acids repeated in tandem, has been called the “Kelch repeats.” The BTB/POZ domain has been proposed to function as a protein-protein interaction interface, which organizes higher order structures involved in chromatin folding or cytoskeleton organization (11).
The Kelch-related proteins are a superfamily of proteins conserved in a wide range of organisms, from viruses to mammals. At least 60 Kelch-related proteins have been identified, but their physiological and biochemical functions remain largely uncharacterized (12, 13). The Drosophila Kelch proteins colocalize with actin filaments in a structure called the ring canal, which bridges 15 nurse cells and the oocyte. Drosophila Kelch protein plays an important role in maintaining actin organization during the development of ring canals (6, 8). The Kelch-related proteins have diverse functions in cell morphology, cell organization, and gene expression, and function in multiprotein complexes through contact sites in their β-propeller domains (14). Recently, a new member of the BTB/Kelch repeat family, gigaxonin, was reported to be a pathological target for neurodegenerative disorders in which alterations were found to contain multiple mutations in the Kelch repeats in the neurofilament network (15).
We have previously identified and characterized two actinbinding proteins, termed NRP/B/ENC-1 (16-18) and Mayven (19), predominantly expressed in brain. Mayven is an actin-binding protein that is co-localized with actin filaments in stress fibers and in the patchy cortical actin-rich regions of the cell margins and processes, including the process tips in primary neurons and U373-MG astrocytoma/glioblastoma cells (19). During our study of proteins that are related to Mayven, we identified and cloned a novel gene, which we termed: MRP2 (Mayven-related protein 2) that was found to be identical to KLHL1 (20, 21). In this study, we have investigated the expression of MRP2/KLHL1 in OLGs and its possible role in the dynamics of cytoskeletal rearrangement, leading to the elongation of OLG processes.

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