Details

Autor(en) / Beteiligte

• Arar, C
• Mignon, C
• Mattei, M
• Monsigny, M
• Roche, A
• Legrand, A

Titel

Mapping of the MR60/ERGIC-53 gene to human chromosome 18q21.3-18q22 by in situ hybridization

Ist Teil von

• Mammalian genome, 1996-10, Vol.7 (10), p.791-792

Ort / Verlag

United States: Springer Verlag

Erscheinungsjahr

1996

Quelle

SpringerLINK Contemporary (Konsortium Baden-Württemberg)

Beschreibungen/Notizen

• Species: Human. Locus name: Intracellular mannose specific lectin. Locus symbol: MR60/ERGIC-53. Map position: 18q21.3-18q22. Method of mapping: in situ hybridization. Database deposit information: GDB: 1220150. Molecular reagents used for mapping: A 4-kb human cDNA probe, isolated from a HL60 cDNA library and inserted into PCRII vector was tritium labeled by nick translation to a specific activity of 1.6 MBq/ mu g. Chromosomes were prepared from lymphoblasts obtained by culturing human lymphocytes for 72 h in the presence of phytohemagglutinin. 5-Bromodeoxyuridine (60 mu g/ml) was added during the last 7 h of culture. The radiolabeled probe was hybridized to metaphase spreads at a final concentration of 25 ng/ml of hybridization solution as described by Mattei and associates. After being coated with nuclear track emulsion (Kodak NTB sub(2)), the slides were exposed for 20 days at 4 degree C, then developed. To avoid any slipping of silver grains during the banding procedure, chromosome spreads were first stained with buffered Giemsa solution and metaphases were photographed. R-banding was then performed by the fluorochrome-photolysis-Giemsa method, and metaphases were rephotographed before analysis. Outstanding conclusions or implications: MR60 is a membrane mannose-specific lectin identified in intracellular compartments of HL60 cells. MR60 represents a new type of animal membrane lectin not related to any known mammalian lectin, although it shares structural homologies with galectins, beta -galactoside-specific animal lectins, and with leguminous lectins. MR60 appeared to be identical to ERGIC-53, a protein marker of the intermediate compartment shuttling between the endoplasmic reticulum and the cis-Golgi apparatus. This protein is putatively involved in the transport of early processed glycoproteins containing high mannose glycans from the ER to the Golgi. In this paper, we map the human MR60/ERGIC-53 gene by in situ hybridization to the 18q 21.3-18q 22 region of the human Chromosome (Chr) 18. On the 100 metaphase cells examined upon in situ hybridization, there were 152 silver grains associated with chromosomes; among them, 69 (45.4%) were located on Chr 18; the distribution of grains on this chromosome was not random: 85% (59/69) of them mapped to the q21.3-q22 region of Chr 18. These results unequivocally map MR60 specific probe to the 18q21.3-18q22 region of the human genome. To date, MR60/ERGIC-53 gene is the first lectin to be located in the Chr 18. All the other known lectin genes were mapped in various other chromosomes. The MR60/ERGIC-53 gene locus is located close to the c-yes-1 and bcl-2 oncogenes, which correspond to a t(14; 18) chromosomal translocation in a follicular lymphoma cell line. A region deleted in a majority of colorectal carcinoma is also located close to that of MR60 /ERGIC-53 gene.