Molecular cloning and characterization of a novel V-ATPase associated protein, DVA9.2, from human dendritic cells.
The vacuolar proton-ATPase (V-ATPase) is a ubiquitous ATP-driven H(+) transporter that functions in numerous cell processes. Accumulating evidence shows important roles of V-ATPase in tumor metastasis and antigen presentation of dendritic cells (DC). A novel V-ATPase associated protein, designated as DVA9.2 (dendritic cell-derived V-ATPase associated protein of 9.2 kDa), has been identified from a human DC cDNA library by large-scale random sequencing. Full length cDNA of DVA9.2 encodes an 81-residue protein that shares 70-80% homology with human V-ATPase subunit M9.2. Distant relationship is also found with Vma21p, a yeast protein required for V-ATPase assembly. DVA9.2 contains a conserved domain, ATP synthase subunit H (pafm05493), and two membrane-spanning helices. DVA9.2 mRNA is detectable in several human tumor cell lines as well as some human normal cells and tissues. Moreover, the inducible expression of DVA9.2 mRNA in DC during maturation is observed. DVA9.2 displays integration with membrane and main localization in lysosome, endoplasmic reticulum and Golgi-associated organelles, only less at the plasma membrane. In addition, DVA9.2 is co-localized with V(0)-sector subunit a. Silencing of DVA9.2 by small interfering RNA (siRNA) does not affect the V-ATPase activity in cell membrane fractions or attenuate the migration and invasion in breast cancer MDA-MB-231 cells. These results indicate that DVA9.2, as a novel V-ATPase-associated protein, is not essential for the activity of V-ATPase complex and may be involved in functions of DC.
Liu
Xingguang
X
Institute of Immunology, Second Military Medical University, Shanghai, PR China.
Wang
Chunmei
C
Li
Nan
N
Zhang
Xuan
X
Zheng
Yuanyuan
Y
Cao
Xuetao
X
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002465:Cell Movement; D022082:Cellular Structures / Q000201:enzymology / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D003713:Dendritic Cells / Q000201:enzymology / Q000378:metabolism / Q000502:physiology; D020868:Gene Silencing; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism / Q000502:physiology
D018076:DNA, Complementary; D012333:RNA, Messenger; D025262:Vacuolar Proton-Translocating ATPases