SUMMARY PARAGRAPH for PAM18
Hsp40/DnaJ is a family of proteins, established by bacterial DnaJ, that regulates Hsp70 chaperone activity. Hsp40s stimulate the intrinsically weak ATPase activity of Hsp70 proteins and facilitate Hsp70 interaction with polypeptide substrates. Hsp70 family members often have multiple Hsp40 partners, and these specific pairings govern Hsp70 chaperone involvement in particular processes (reviewed in 5, 6, and 7). All Hsp40s contain a highly conserved 75-amino acid J domain, which interacts with the ATPase domain of Hsp70 to stimulate ATP hydrolysis. However, there are also other conserved structural domains, and based on the presence or absence of these regions, the Hsp40 family can be divided into three subtypes: type I, type II and type III (a comprehensive overview of the structural features of the different HSP40 subtypes can be found in 7). Sequence analysis of the S. cerevisiae genome has revealed 22 proteins in the Hsp40/DnaJ family: YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2, JJJ1, JJJ2, JJJ3, CAJ1, CWC23, MDJ1, MDJ2, PAM18, JAC1, JID1, SCJ1, HLJ1, JEM1, SEC63, and ERJ5 (7).
About mitochondrial import
While the mitochondrial genome encodes a handful of proteins, most of the hundreds of proteins that reside in the mitochondrion are encoded by nuclear genes, translated in the cytoplasm, and imported into mitochondria via a series of complex molecular machines (see 8, 9 for review). Many of the proteins imported into mitochondria are involved in respiration, which is not an essential process: S. cerevisiae is able to carry out either fermentative growth on carbon sources such as glucose, or respiratory growth on nonfermentable carbon sources such as glycerol and ethanol. However, since maintenance of the mitochondrial compartment is essential to life, mutations that completely disrupt mitochondrial import are lethal.
About the TIM23 complex
The Translocase of the Inner Mitochondrial membrane (TIM23 complex) receives proteins from the Translocase of the Outer Mitochondrial membrane (TOM complex) and either directs them into the mitochondrial matrix or facilitates their integration into the mitochondrial inner membrane (reviewed in 10, 9, 11). The membrane-embedded core of the complex is composed of three essential proteins: Tim23p, Tim17p, and Tim50p. Tim23p and Tim17p, which share sequence similarity, comprise the twin-pore structure through which precursor proteins translocate. Tim23p alone has the ability to form a voltage-sensitive channel (12), but Tim17p is required in vivo for maintenance of the twin-pore architecture and for normal function of the pore (13). Tim17p also has a role in sorting incoming proteins to the mitochondrial matrix or the inner membrane (14). Tim50p interacts with precursor proteins and with Tim23p to guide precursors from the TOM complex to the TIM23 complex (15, 16). Two additional non-essential components, Tim21p and Pam17p, interact with the core of the TIM23 complex and may modulate its activity (14, 17, 18).
Proteins destined for the mitochondrial matrix require the action of a sub-complex of the TIM23 complex, known as the import motor or presequence translocase-associated motor (PAM) complex. Its catalytic component is Ssc1p, a member of the heat shock 70 protein family commonly referred to as mtHsp70, which undergoes cycles of binding and release of the precursor, hydrolyzing ATP and changing conformation in the process. The nucleotide release factor Mge1p promotes this cycle by facilitating the dissociation of ADP from Ssc1p (19, 20). Other components include Tim44p, an essential subunit that mediates the association of the core TIM23 complex with the PAM complex (21, 18); Pam18p (Tim14p), a J-protein cochaperone that stimulates the ATPase activity of Ssc1p; and Pam16p (Tim16p), a J-like protein that binds to Pam18p and regulates its activity (4). Pam17p mediates the association between Pam16p and Pam18p (22). Once imported proteins reach the mitochondrial matrix, their correct folding is facilitated by a soluble complex consisting of Ssc1p and its cochaperones Mdj1p and Mge1p (23).
A subset of proteins destined for insertion into the mitochondrial inner membrane is translocated via the TIM23 complex but then inserted laterally into the inner membrane rather than entering the mitochondrial matrix. This mechanism is currently not understood in detail. The TIM23 complex adopts different conformations during the two kinds of import, but it is unclear whether this inner membrane import is accomplished by the core complex alone (Tim23p, Tim17p, and Tim50p), or by the entire TIM23 complex including the import motor subunits (10, 17).
About PAM16, PAM17, and PAM18
Pam16p and Pam18p are both related to J-proteins, which act as cochaperones for chaperone proteins belonging to the Hsp70 family. Pam18p is considered a J-protein because it has all of the characteristic sequence elements, while Pam16p is "J-like" because it lacks the conserved HPD motif (24, 25, 1). Both proteins are constituents of the import motor (PAM) complex, and interact with each other to form a heterodimer (26, 4). Pam18p stimulates the ATPase activity of Ssc1p, which is the catalytic component of the import motor, while Pam16p inhibits the stimulatory activity of Pam18p (27, 26).
The localization and role of Pam17p are currently unclear. It is a conserved protein that was identified as a component of the import motor complex (22), but other studies have detected interactions between Pam17p and components of the TIM23 core complex, and not between Pam17p and the import motor subunits (17). It has been proposed that Pam17p mediates association of the Pam16p-Pam18p heterodimer with the TIM23 complex, and alternatively that its role is to modulate the function of the core complex by affecting its conformation (17, 22).
In keeping with their proposed roles in the import motor, mutations in all three of the PAM genes affect the ability of the TIM23 complex to import proteins into the mitochondrial matrix, but not its ability to insert proteins into the inner membrane (24, 25, 28, 22). PAM16 and PAM18 are essential genes (26, 4), while a pam17 null mutant is viable but displays a slow respiratory growth phenotype (22).
Last updated: 2006-12-19