title: A unique carboxyl-terminal insert domain in the hematopoietic-specific, GTPase-deficient Rho GTPase RhoH regulates post-translational processing.
abstract: RhoH is a hematopoietic-specific, GTPase-deficient member of the Rho GTPase family that was first identified as a hypermutable gene in human B lineage lymphomas. RhoH remains in a constitutively active state and thus its effects are regulated by expression levels or post-translational modifications. Similar to other small GTPases, intracellular localization of RhoH is dependent upon the conserved "CAAX" box and surrounding sequences within the carboxyl (C) terminus. However, RhoH also contains a unique C-terminal "insert" domain of yet undetermined function. RhoH serves as adaptor molecule in T cell receptor signaling and RhoH expression correlates with the unfavorable prognostic marker ZAP70 in human chronic lymphocytic leukemia. Disease progression is attenuated in a Rhoh(-/-) mouse model of chronic lymphocytic leukemia and treatment of primary human chronic lymphocytic leukemia cells with Lenalidomide results in reduced RhoH protein levels. Thus, RhoH is a potential therapeutic target in B cell malignancies. In the current studies, we demonstrate that deletion of the insert domain (LFSINE) results in significant cytoplasmic protein accumulation. Using inhibitors of degradation pathways, we show that LFSINE regulates lysosomal RhoH uptake and degradation via chaperone-mediated autophagy. Whereas the C-terminal prenylation site is critical for ZAP70 interaction, subcellular localization and rescue of the Rhoh(-/-) T cell defect in vivo, the insert domain appears dispensable for these functions. Taken together, our findings suggest that the insert domain regulates protein stability and activity without otherwise affecting RhoH function.
authors.affiliation: From the Division of Hematology/Oncology, Boston Children's Hospital and the Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
authors.forename: Anja
authors.initials: A
authors.lastname: Troeger
authors.forename: Hee-Don
authors.initials: HD
authors.lastname: Chae
authors.forename: Mumine
authors.initials: M
authors.lastname: Senturk
authors.forename: Jenna
authors.initials: J
authors.lastname: Wood
authors.forename: David A
authors.initials: DA
authors.lastname: Williams
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D054876:Prenylation; D011499:Protein Processing, Post-Translational; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D059748:Proteolysis; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C496930:RhoH protein, mouse; D014157:Transcription Factors; D020741:rho GTP-Binding Proteins
keywords: Atypical Rho GTPases; Autophagy; B-Cell Malignancies; Carboxyl Terminus; Chaperone-mediated Autophagy; Protein Degradation; Protein Stability; Rho GTPases; RhoH; T Cell