Role of the Lysosomal Membrane Protein, CLN3, in the Regulation of Cathepsin D Activity

Among Neuronal Ceroid Lipofuscinoses (NCLs), which are childhood fatal neurodegenerative disorders, the juvenile onset form (JNCL) is the most common. JNCL is caused by recessive mutations in the CLN3 gene. CLN3 encodes a lysosomal/endosomal transmembrane protein but its precise function is not completely known. We have previously reported that in baby hamster kidney (BHK) cells stably expressing myc‐tagged human CLN3 (myc‐CLN3), hyperosmotic conditions drastically increased myc‐CLN3 mRNA and protein expression. In the present study, we analyzed the consequences of hyperosmolarity, and increased CLN3 expression on cathepsin D (CTSD) activity and prosaposin processing using BHK cells transiently or stably expressing myc‐CLN3. We found that hyperosmolarity increased lysotracker staining of lysosomes, and elevated the levels of myc‐CLN3 and lysosome‐associated membrane protein‐1 (LAMP1). Hyperosmolarity, independently of the expression level of myc‐CLN3, decreased the levels of PSAP and saposin D, which are protein cofactors in sphingolipid metabolism. The lysosomal enzyme cathepsin D (CTSD) mediates the proteolytic cleavage of PSAP precursor into saposins A‐D. Myc‐CLN3 colocalized with CTSD and activity of CTSD decreased as myc‐CLN3 expression increased, and clearly decreased under hyperosmotic conditions. Nevertheless, levels of CTSD measured by Western blotting were not altered under any studied condition. Our results suggest a direct involvement of CLN3 in the regulation of CTSD activity. J. Cell. Biochem. 118: 3883–3890, 2017. © 2017 Wiley Periodicals, Inc.

[1]  M. Raad,et al.  Association between CLN3 (Neuronal Ceroid Lipofuscinosis, CLN3 Type) Gene Expression and Clinical Characteristics of Breast Cancer Patients , 2015, Front. Oncol..

[2]  Jian Dong,et al.  Activation of autophagy via Ca2+-dependent AMPK/mTOR pathway in rat notochordal cells is a cellular adaptation under hyperosmotic stress , 2015, Cell cycle.

[3]  Michael A. Myre,et al.  Loss of Cln3 Function in the Social Amoeba Dictyostelium discoideum Causes Pleiotropic Effects That Are Rescued by Human CLN3 , 2014, PloS one.

[4]  A. Petcherski,et al.  FRET-Assisted Determination of CLN3 Membrane Topology , 2014, PloS one.

[5]  J. Matsuda,et al.  Mesotrypsin and Caspase-14 Participate in Prosaposin Processing , 2014, The Journal of Biological Chemistry.

[6]  Larissa A. Haliw,et al.  Human iPSC models of neuronal ceroid lipofuscinosis capture distinct effects of TPP1 and CLN3 mutations on the endocytic pathway. , 2014, Human molecular genetics.

[7]  A. Jalanko,et al.  Cell biology and function of neuronal ceroid lipofuscinosis-related proteins. , 2013, Biochimica et biophysica acta.

[8]  D. Pearce,et al.  Osmotic Stress Changes the Expression and Subcellular Localization of the Batten Disease Protein CLN3 , 2013, PloS one.

[9]  Sung-Jo Kim,et al.  Cell cycle arrest in Batten disease lymphoblast cells. , 2013, Gene.

[10]  E. Knecht,et al.  Alterations in ROS Activity and Lysosomal pH Account for Distinct Patterns of Macroautophagy in LINCL and JNCL Fibroblasts , 2013, PloS one.

[11]  Angelika Saje,et al.  Age-dependent therapeutic effect of memantine in a mouse model of juvenile Batten disease , 2012, Neuropharmacology.

[12]  S. Mole,et al.  New nomenclature and classification scheme for the neuronal ceroid lipofuscinoses , 2012, Neurology.

[13]  S. Cotman,et al.  The juvenile Batten disease protein, CLN3, and its role in regulating anterograde and retrograde post-Golgi trafficking , 2012, Clinical lipidology.

[14]  J. Neefjes,et al.  Neuronal ceroid lipofuscinosis protein CLN3 interacts with motor proteins and modifies location of late endosomal compartments , 2012, Cellular and Molecular Life Sciences.

[15]  C. A. Beck,et al.  Quantifying physical decline in juvenile neuronal ceroid lipofuscinosis (Batten disease) , 2011, Neurology.

[16]  I. Martins,et al.  Osmoregulation of ceroid neuronal lipofuscinosis type 3 in the renal medulla. , 2010, American journal of physiology. Cell physiology.

[17]  L. Ricci-Vitiani,et al.  The secretion and maturation of prosaposin and procathepsin D are blocked in embryonic neural progenitor cells. , 2008, Biochimica et biophysica acta.

[18]  R. Boustany,et al.  CLN3p Impacts Galactosylceramide Transport, Raft Morphology, and Lipid Content , 2008, Pediatric Research.

[19]  Yusuf A. Hannun,et al.  Principles of bioactive lipid signalling: lessons from sphingolipids , 2008, Nature Reviews Molecular Cell Biology.

[20]  K. Roth,et al.  The autophagy-lysosomal degradation pathway: role in neurodegenerative disease and therapy. , 2008, Frontiers in bioscience : a journal and virtual library.

[21]  A. Lehesjoki,et al.  Cathepsin D deficiency underlies congenital human neuronal ceroid-lipofuscinosis. , 2006, Brain : a journal of neurology.

[22]  G. Guidotti,et al.  Amino acid signaling through the mammalian target of rapamycin (mTOR) pathway: Role of glutamine and of cell shrinkage , 2005, Journal of cellular physiology.

[23]  Y. Hannun,et al.  Updates on functions of ceramide in chemotherapy-induced cell death and in multidrug resistance. , 2001, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[24]  J. Matsuda,et al.  A mutation in the saposin A domain of the sphingolipid activator protein (prosaposin) gene results in a late-onset, chronic form of globoid cell leukodystrophy in the mouse. , 2001, Human molecular genetics.

[25]  F. Sherman,et al.  The Yeast Model for Batten Disease: Mutations inbtn1, btn2, and hsp30 Alter pH Homeostasis , 2000, Journal of bacteriology.

[26]  K. Wisniewski,et al.  CLN3 protein regulates lysosomal pH and alters intracellular processing of Alzheimer's amyloid-beta protein precursor and cathepsin D in human cells. , 2000, Molecular genetics and metabolism.

[27]  H. Nakanishi,et al.  Cathepsin D deficiency induces lysosomal storage with ceroid lipofuscin in mouse CNS neurons. , 2000, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[28]  W. Schneider-Brachert,et al.  Cathepsin D targeted by acid sphingomyelinase‐derived ceramide , 1999, The EMBO journal.

[29]  T. Ferea,et al.  Action of BTN1, the yeast orthologue of the gene mutated in Batten disease , 1999, Nature Genetics.

[30]  B. Martin,et al.  Lysosomal proteolysis of prosaposin, the precursor of saposins (sphingolipid activator proteins): its mechanism and inhibition by ganglioside. , 1997, Archives of biochemistry and biophysics.

[31]  J. Haines,et al.  Isolation of a novel gene underlying batten disease, CLN3 , 1995, Cell.

[32]  N. Rawlings,et al.  Families of aspartic peptidases, and those of unknown catalytic mechanism. , 1995, Methods in enzymology.

[33]  M. Horowitz,et al.  Mutations causing gaucher disease , 1994, Human mutation.

[34]  M. Baumann,et al.  Storage of saposins A and D in infantile neuronal ceroid‐lipofuscinosis , 1993, FEBS letters.

[35]  Y. Kishimoto,et al.  Saposins: structure, function, distribution, and molecular genetics. , 1992, Journal of lipid research.

[36]  Y. Moriyama,et al.  Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. , 1991, The Journal of biological chemistry.

[37]  M. Fukuda,et al.  Structure of human lysosomal membrane glycoprotein 1. Assignment of disulfide bonds and visualization of its domain arrangement. , 1989, The Journal of biological chemistry.