Integrin and spectrin homologues, and cytoplasm-wall adhesion in tip growth.
Saprolegnia ferax contains an integrin homologue, identified by crossreactivity with antiserum to the consensus sequence of human/chick/Xenopus cytoplasmic domain beta 1-integrin, which is highly conserved. In non-reduced samples, this integrin was larger than the reported size range for beta 1-integrins, at 178 kDa. In reduced samples, there was a reducing agent-concentration-dependent conversion from 178 kDa to 120 kDa, well within the reported size range for beta 1-integrins in other organisms. The integrin antiserum stained plasma membrane-associated patches, which had a shallow tip-high gradient. This population was reduced and its distribution perturbed in hyphae whose growth rate was reduced by half with tetrapentyl ammonium chloride. The expected integrin function in cytoplasm-cell wall attachment was shown by differential resistance to plasmolysis-induced separation, which positively correlated with integrin abundance. However, when there was separation, remnants of cytoplasm stayed attached to the wall. These were enriched in actin and integrin. Saprolegnia also has a spectrin homologue identified by crossreactivity with an erythrocyte spectin antibody, which has a size (246 kDa) similar to other organisms. This spectrin had a superficially similar distribution to that of integrin, but it did not participate in cytoplasm-wall anchoring. These data suggest that Saprolegnia hyphae have a plasma membrane which is strengthened by spectrin, and cytoplasm which is attached to the cell wall by integrin.
Kaminskyj
S G
SG
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392, USA.
Heath
I B
IB
D000818:Animals; D015153:Blotting, Western; D002473:Cell Wall / Q000378:metabolism*; D002645:Chickens; D003593:Cytoplasm / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism* / Q000502:physiology; D009868:Oomycetes / Q000254:growth & development / Q000378:metabolism*; D013049:Spectrin / Q000378:metabolism* / Q000502:physiology; D014981:Xenopus
D016023:Integrins; D013049:Spectrin