OBJECTIVETo study the characteristics of a novel human testis-specific gene, HSD-9, and its encoding protein.

METHODSHSD-9 was a novel gene from a human germ cells-specific ESTs library established in our laboratory. We used an electronic cloning method to obtain HSD-9 gene, and analyzed the characteristics of this novel gene and encoding product by bioinformatics methods, detected its expressing profile using a Northern blot assay, prepared specific rabbit polyclonal antibodies against HSD-9 protein, observed the localization of this protein in the germ cells and some somatic cells with confocal microscopy.

RESULTSHSD-9 was expressed in human testes, and its rat homolog was found in the varying germ cells. HSD-9 protein could partly be colocalized with clathrin.

CONCLUSIONSHSD-9 is specific in human testes, and the expression pattern of its encoding product is similar to those of some endocytosis proteins. It is speculated that HSD-9 protein may function in the endocytosis.

Amino Acid Sequence ; Animals ; Base Sequence ; Clathrin ; metabolism ; Humans ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Organ Specificity ; Rabbits ; Rats ; Testis ; metabolism

Aanaplastic lymphoma kinase (ALK)-positive diffuse large B-cell lymphoma (DLBCL) is an unusual disease entity first reported in 1997 as DLBCL with expression of full-length ALK protein. The World Health Organization classification enlists the disease as a rare variant of DLBCL. Herein we describe two cases of ALK-positive DLBCL with cytomorphologic and molecular characteristics for the first time in Korea. The patients were 35-yr-old and 24-yr-old male patients. Immunohistochemical studies on the lymph nodes revealed large sized neoplastic cells with plasmablastic differentiation, which were negative for CD30 and positive for ALK with the characteristic granular staining in the cytoplasmic region. Extensive involvement of bone marrow was observed in both cases showing large, extremely atypical cells. Fluorescence in situ hybridization and molecular studies on the bone marrow aspirate specimens led to the detection of a clathrin (CLTC)/ALK rearrangement. Despite aggressive chemotherapy, the patients died 15 and 17 months after the diagnosis, indicating poor prognosis of the disease entity. This is the first report demonstrating the cytomorphologic findings of ALK-positive DLBCL cells on bone marrow aspirates.
Adult ; Bone Marrow/*pathology ; Clathrin/*genetics ; Fatal Outcome ; *Gene Fusion ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse/*genetics/metabolism/*pathology ; Male ; Protein-Tyrosine Kinases/*genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

The translocation t(10;11)(p13;q14q21) has been found to be recurrent in acute lymphoblastic and myeloid leukemias, and results in the fusion of the clathrin assembly lymphoid myeloid leukemia (CALM) gene with the AF10 gene; these genes are present on chromosomes 11 and 10, respectively. Because the CALM-AF10 rearrangement is a rare chromosomal abnormality, it is not included in routine molecular tests for acute leukemia. Here, we describe the cases of 2 patients with the CALM-AF10 fusion gene. The first patient (case 1) was diagnosed with T-cell ALL, and the second patient (case 2) was diagnosed with AML. Both patient samples showed expression of the homeobox A gene cluster and the histone methyltransferase hDOT1L, which suggests that they mediate leukemic transformation in CALM-AF10-positive and mixed-lineage leukemia-AF10-positive leukemias. Both patients achieved complete remission after induction chemotherapy. The first patient (case 1) relapsed after double-unit cord blood transplantation; there was no evidence of relapse in the second patient (case 2) after allogenic peripheral blood stem cell transplantation. Since CALM-AF10- positive leukemias have been shown to have poor prognosis with conventional therapy, molecular tests for CALM-AF10 rearrangement would be necessary to detect minimal residual disease during follow-up.
Adolescent ; Adult ; Bone Marrow/pathology ; Chromosomes, Human, Pair 10 ; Chromosomes, Human, Pair 11 ; Cord Blood Stem Cell Transplantation ; Female ; Histone-Lysine N-Methyltransferase/genetics/metabolism ; Homeodomain Proteins/genetics/metabolism ; Humans ; Leukemia, Myeloid, Acute/diagnosis/*genetics/therapy ; Male ; Monomeric Clathrin Assembly Proteins/*genetics ; Oncogene Proteins, Fusion/*genetics ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/diagnosis/*genetics/therapy ; Recurrence ; Transcription Factors/*genetics ; Translocation, Genetic

Clathrin-coated vesicles (CCVs) are involved in protein and lipid trafficking between intracellular compartments in eukaryotic cells. CCVs are composed of clathrin and assembly proteins. The clathrin assembly protein lymphoid myeloid leukemia (CALM) gene, encodes a homologoue of the neuronal clathrin assembly protein AP180. In this study, we characterized the properties of the CALM expressed in E. coli. The molecular weight of bacterially expressed GST-CALM fusion protein was approximately 105 kD on SDS-PAGE. The CALM protein could promote clathrin triskelia into clathrin cages and could bind the preformed clathrin cage. However, 33 kD N-terminal domain of CALM could not bind pre-assembled clathrin cages, but assemble clathrin triskelia into clathrin cages. The CALM protein was bound to SH3 domain through N-terminal domain1, in vitro. The CALM protein is proteolyzed by caspase 3, caspase 8 and calpain through C-terminal domain.
Animal ; Antibodies, Monoclonal ; Calpain/chemistry ; Caspases/chemistry ; Clathrin-Coated Vesicles/metabolism* ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli/metabolism ; Escherichia coli/genetics ; Female ; Glutathione Transferase/genetics* ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/chemistry* ; Phosphoproteins/metabolism ; Phosphoproteins/genetics ; Phosphoproteins/chemistry* ; Protein Binding ; Rabbits ; Recombinant Fusion Proteins/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/chemistry* ; src Homology Domains