title: Hyaluronidase Hyal1 Increases Tumor Cell Proliferation and Motility through Accelerated Vesicle Trafficking.
abstract: Hyaluronan (HA) turnover accelerates metastatic progression of prostate cancer in part by increasing rates of tumor cell proliferation and motility. To determine the mechanism, we overexpressed hyaluronidase 1 (Hyal1) as a fluorescent fusion protein and examined its impact on endocytosis and vesicular trafficking. Overexpression of Hyal1 led to increased rates of internalization of HA and the endocytic recycling marker transferrin. Live imaging of Hyal1, sucrose gradient centrifugation, and specific colocalization of Rab GTPases defined the subcellular distribution of Hyal1 as early and late endosomes, lysosomes, and recycling vesicles. Manipulation of vesicular trafficking by chemical inhibitors or with constitutively active and dominant negative Rab expression constructs caused atypical localization of Hyal1. Using the catalytically inactive point mutant Hyal1-E131Q, we found that enzymatic activity of Hyal1 was necessary for normal localization within the cell as Hyal1-E131Q was mainly detected within the endoplasmic reticulum. Expression of a HA-binding point mutant, Hyal1-Y202F, revealed that secretion of Hyal1 and concurrent reuptake from the extracellular space are critical for rapid HA internalization and cell proliferation. Overall, excess Hyal1 secretion accelerates endocytic vesicle trafficking in a substrate-dependent manner, promoting aggressive tumor cell behavior.
authors: From the Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588.
authors: Caitlin O
authors: CO
authors: McAtee
authors: From the Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588.
authors: Abigail R
authors: AR
authors: Berkebile
authors: Morrison Microscopy Facility, University of Nebraska, Lincoln, Nebraska 68588.
authors: Christian G
authors: CG
authors: Elowsky
authors: Morrison Microscopy Facility, University of Nebraska, Lincoln, Nebraska 68588.
authors: Teresa
authors: T
authors: Fangman
authors: From the Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588.
authors: Joseph J
authors: JJ
authors: Barycki
authors: Department of Oral Biology, University of Nebraska Medical Center College of Dentistry, Lincoln, Nebraska 68503.
authors: James K
authors: JK
authors: Wahl
authors: From the Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588.
authors: Oleh
authors: O
authors: Khalimonchuk
authors: Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, Nebraska 68198, and.
authors: Naava
authors: N
authors: Naslavsky
authors: Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, Nebraska 68198, and Fred and Pamela Buffett Cancer Center, Omaha, Nebraska 68198.
authors: Steve
authors: S
authors: Caplan
authors: From the Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588, Fred and Pamela Buffett Cancer Center, Omaha, Nebraska 68198 msimpson2@unl.edu.
authors: Melanie A
authors: MA
authors: Simpson
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology; D000951:Antigens, Neoplasm / Q000378:metabolism*; D017209:Apoptosis; D015153:Blotting, Western; D002465:Cell Movement; D049109:Cell Proliferation; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism*; D051548:Histone Acetyltransferases / Q000378:metabolism*; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D008297:Male; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D021381:Protein Transport; D013347:Subcellular Fractions; D014168:Transferrin / Q000378:metabolism; D022161:Transport Vesicles / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D014168:Transferrin; D006820:Hyaluronic Acid; D051548:Histone Acetyltransferases; C115866:OGA protein, human; D006821:Hyaluronoglucosaminidase
keywords: cell motility; endocytosis; hyaluronan; hyaluronidase; lysosome; prostate cancer